Elimination of hepatitis b virus with antiviral agents

ABSTRACT

The present invention is directed to compounds, compositions and methods for preventing, treating or curing Hepatitis B (HBV) infection in human subjects or other animal hosts. The compounds are as also pharmaceutically acceptable, salts, prodrugs, and other derivatives thereof as pharmaceutical compositions and methods for treatment, prevention or eradication of HBV infection.

FIELD OF THE INVENTION

The present invention is directed to compounds, methods and compositionsfor preventing, treating and/or curing hepatitis B virus (HBV)infections. More specifically, the invention describes specificallysubstituted aromatic/heteroaromatic compounds, pharmaceuticallyacceptable salts, or other derivatives thereof, and the use thereof inthe treatment of HBV infections.

BACKGROUND OF THE INVENTION

Hepatitis B virus (HBV) causes a serious human health problem and issecond only to tobacco as a cause of human cancer. The mechanism bywhich HBV induces cancer is unknown. It is postulated that it maydirectly trigger tumor development, or indirectly trigger tumordevelopment through chronic inflammation, cirrhosis, and cellregeneration associated with the infection.

After a 2- to 6-month incubation period, during which the host istypically unaware of the infection, HBV infection can lead to acutehepatitis and liver damage, resulting in abdominal pain, jaundice andelevated blood levels of certain enzymes. HBV can cause fulminanthepatitis, a rapidly progressive, often fatal form of the disease inwhich large sections of the liver are destroyed.

Subjects typically recover from the acute phase of HBV infection. Insome patients, however, the virus continues replication for an extendedor indefinite period, causing a chronic infection. Chronic infectionscan lead to chronic persistent hepatitis. Patients infected with chronicpersistent HBV are most common in developing countries. By mid-1991,there were approximately 225 million chronic carriers of HBV in Asiaalone and worldwide almost 300 million carriers. Chronic persistenthepatitis can cause fatigue, cirrhosis of the liver, and hepatocellularcarcinoma, a primary liver cancer.

In industrialized countries, the high-risk group for HBV infectionincludes those in contact with HBV carriers or their blood samples. Theepidemiology of HBV is very similar to that of HIV/AIDS, which is areason why HBV infection is common among patients infected with HIV orsuffering from AIDS. However, HBV is more contagious than HIV.

3TC (lamivudine), interferon alpha-2b, peginterferon alpha-2a, hepsera(adefovir dipivoxil), baraclude (entecavir), and Tyzeka (Telbivudine)are currently FDA-approved drugs for treating HBV infection. Anothernucleoside, tenofovir alafenamide fumarate (TAF) (formerly GS-7340) iscurrently in phase 3. All these drugs are highly effective in reducingviral load, but none of these drugs provide a cure for HBV. In addition,their impact can be limited by drug resistance, low efficacy andtolerability issues. The low cure rates of HBV are attributed at leastin part to the presence and persistence of covalently closed circularDNA (cccDNA) in the nucleus of infected hepatocytes.

Accordingly, there is an urgent need for new HBV drugs that are potent,safe, that work by a different mechanism than nucleoside analogs, andcan reduce the latent form of HBV known as cccDNA.

It would be advantageous to provide new antiviral agents, compositionsincluding these agents, and methods of treatment using these agents totreat HBV and prevent the emergence of drug-resistant HBV. The presentinvention provides such agents, compositions and methods.

SUMMARY OF THE INVENTION

The present invention provides compounds, methods and compositions forpreventing, treating and/or curing HBV infection in a host, or reducingthe activity of HBV in the host. The methods involve administering atherapeutically or prophylactically-effective amount of at least onecompound as described herein to treat, cure or prevent an infection by,or an amount sufficient to reduce the biological activity of, an HBVinfection.

The compounds can also be used to treat other viral infections,including those by flaviviridae viruses, such as West Nile virus (WNV)and, hepatitis C virus (HCV), Dengue Fever, Zika virus.

The pharmaceutical compositions include one or more of the compoundsdescribed herein, in combination with a pharmaceutically acceptablecarrier or excipient, for treating a host infected with HBV. Thesecompounds can be used in combination with nucleoside and non-nucleosideinhibitors of HBV. The formulations can further include at least oneother therapeutic agent. In addition, the present invention includesprocesses for preparing such compounds.

In one embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof,

wherein:

A is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl;

B is a six or seven-membered ring or a six or seven-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S, a five-membered ring containing zero, one, ortwo heteroatoms, which are, independently, N, O, or S; a four-memberedring containing zero, one, or two heteroatoms, which are, independently,N, O, or S, or a C₅₋₁₄ bicyclic ring,

When R¹ and R^(1′) are attached to a carbon, they are, independently,hydrogen, halogen (including F, Cl, Br, and I), SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₂₋₆alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl;

When R¹ and R^(1′) are attached to a nitrogen, they are, independently,hydrogen, C₂₋₆ alkoxy, C₃₋₆ alkoxyalkyl, C₂₋₆ alkenyl, alkoxycarbonyl,carbonylalkyl, carbonyl aryl, C₁₋₆ alkyl, heterocyclylalkyl, C₂₋₆hydroxyalkyl, or S(O)₂R′;

Each R′ is independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl,alkylaryl, or arylalkyl, or if two R′ reside on the same nitrogen atom,they can come together to form a C₃₋₆ ring optionally containing a N, O,or S heteroatom;

The R′ groups can optionally be substituted with one or moresubstituents, which substituents are, independently, halo, C₁₋₆haloalkyl, C₁₋₆ hydroxyalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy,amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, alkoxyalkyl, aryloxy, nitro, cyano, sulfonic acid, thiol, imine,sulfonyl, sulfanyl, sulfinyl, sulfamoyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid, orphosphonate, either unprotected, or protected as necessary, as known tothose skilled in the art, for example, as taught in Greene, et al.,Protective Groups in Organic Synthesis, John Wiley and Sons, SecondEdition, 1991, hereby incorporated by reference;

u and v are independently 0, 1, 2, 3, 4 or 5;

X is

R³ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R² is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, such as phenyl, heteroaryl, including six-memberedheteroaromatic rings containing one, two, or three nitrogen atoms andfive-membered heteroaromatic rings containing one, two, or threeheteroatoms, which, independently, are N, O, or S, alkylaryl, arylalkyl,a six-membered ring or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, a seven-membered bridged or spiro-fused ring containing zero, one,or two heteroatoms which are, independently, N, O, or S, a five-memberedring containing zero, one, or two heteroatoms which are, independently,N, O, or S; a four-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; cycloalkyl,alkylheteroaryl, or alkylaryl;

R² is optionally substituted with one or more substituents, which each,independently, are halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or isoptionally substituted with aryl, substituted aryl, heteroaryl, orsubstituted heteroaryl, where substituents on the substituted aryl andsubstituted heteroaryl are selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl.

or

R² and R³ can come together with the nitrogen to which they are attachedto form a 6-10 membered bicyclic or bridged ring, a 3 to 8 saturatedring, or a 5 membered unsaturated ring; such bicyclic, bridged,saturated and unsaturated rings optionally containing one or moreadditional heteroatoms, where each is, independently, O, S or N, andoptionally being substituted with one or more substituents, whereineach, independently, is halogen (including F, Cl, Br, I), CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl.

In a second embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

C is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylaryl, or alkylheteroaryl;

D is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S, ora C₅₋₁₄ bicyclic ring,

Y is

R⁴ is H or C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl; inone embodiment, R⁴ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl,

R⁵ is alkylaryl, arylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, such asphenyl, heteroaryl, including six-membered heteroaromatic ringscontaining one, two, or three nitrogen atoms and five-memberedheteroaromatic rings containing one, two, or three heteroatoms, which,independently, are N, O, or S; and a six-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S; in one embodiment, R⁵ is alkylaryl, arylalkyl, phenyl, afive or six-membered heteroaryl, or a six-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S;

R⁵ is optionally substituted with one or more substituents, each ofwhich is, independently, halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; or is substituted with aryl, substituted aryl, heteroaryl,or substituted heteroaryl, where substituents on the substituted aryland substituted heteroaryl are selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl;

where, in one embodiment, if C is phenyl, D is not phenyl or a5-membered ring heteroaryl, and in another embodiment, if C is phenyland D is phenyl or a 5-membered ring heteroaryl, then R⁵ is notalkylaryl, alkenyl, or a six-membered bridged ring;

or when Y is

R⁴ and R⁵ together with the nitrogen to which they are attached to forma 3 to 4 membered ring optionally substituted with one or moresubstituents, each of which is, independently, halogen (including F, Cl,Br, I), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, aryl alkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In one embodiment, D is

where R⁶ is H, Cl, F or Br, and R⁷ is H, methyl, F or Cl.

In one aspect of this embodiment, when Y is

R⁵ is not

In another aspect of this embodiment, when R⁴ is ethyl, then R⁵ is not

In a third embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

E is a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms, where each is, independently, N, O, or S; a C₄₋₁₄bicyclic ring, alkylheteroaryl, or alkylaryl;

F is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S, or a C₄₋₁₄ bicyclicring,

Z is

R⁸ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R⁹ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ringor a six-membered bridged or spiro-fused ring containing zero, one, ortwo heteroatoms, which are independently N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms, which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S, or a three membered ring;

R⁹ is optionally substituted with one or more substituents, each ofwhich is independently halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; or is substituted with aryl, substituted aryl, heteroaryl,or substituted heteroaryl, where substituents on the substituted aryland substituted heteroaryl are selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl; or

R⁸ and R⁹ can come together with the nitrogen to which they are attachedto form a 6-10 membered bicyclic or bridged ring or a 3 to 8 saturatedring, such bicyclic, bridged and saturated ring moiety optionallycontaining one or more additional heteroatoms which, independently, areO, S or N and optionally being substituted with one or moresubstituents, each, independently, is halogen (including F, Cl, Br, andI), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In a fourth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

G is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl;

H is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a six-membered non-aromatic ring optionallycontaining one, two, or three heteroatoms, which are, independently, N,O, or S; or a C₄₋₁₄ bicyclic ring;

When R¹ and R^(1′) are attached to a carbon they are, independently,hydrogen, halogen (including F, Cl, Br, and I), CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, C₂₋₆ alkynyl,C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl;

When R¹ and R^(1′) are attached to a nitrogen they are, independently,hydrogen, C₁₋₆ alkoxy, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, carbonylalkyl,carbonyl aryl, C₁₋₆ alkyl, C₂₋₆ alkynyl, C₂₋₆ alkenyl,heterocyclylalkyl, C₁₋₆ hydroxyalkyl, or S(O)₂R′;

Each R′ is independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl,alkylaryl, or arylalkyl, or if two R′ reside on the same nitrogen atomthey can come together to form a C₃₋₆ alkyl ring optionally containing aN, O, or S; wherein the R′ groups can be substituted with one or moresubstituents as defined above, for example, C₁₋₆ hydroxyalkyl,aminoalkyl, and alkoxyalkyl;

u and v are independently 0, 1, 2, 3, 4 or 5;

W is

R¹⁰ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹¹ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, heteroaryl, alkylaryl, arylalkyl, phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, asix-membered ring or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S, a seven-membered bridged or spiro-fused ring containing zero,one, or two heteroatoms, which are, independently, N, O, or S, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S; a four-membered ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S; a three membered ring,alkylheteroaryl, or alkylaryl;

wherein R¹¹ is optionally substituted with one or more substituentsselected from the group consisting of halogen (including F, Cl, Br, andI), SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, C₁₋₆ hydroxyalkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl;

or

R¹⁰ and R¹¹ can come together with the nitrogen to which they areattached to form a 6-10 membered bicyclic or bridged ring or a 3 to 8saturated ring; such bicyclic, bridged or saturated ring moietyoptionally containing one or more additional heteroatoms, which areeach, independently, O, S or N, and optionally substituted with one ormore substituents, each of which is, independently, halogen (includingF, Cl, Br, and I), CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In a fifth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

I is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S, aC₄₋₁₄ bicyclic ring; alkylheteroaryl, or alkylaryl;

J is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; or a four-memberedring containing zero, one, or two heteroatoms which are, independently,N, O, or S,

W is

R¹² is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹³ is C₂₋₆ alkenyl, C₁₋₆ alkynyl, aryl, including phenyl, heteroaryl,including six-membered heteroaromatic rings containing one, two, orthree nitrogen atoms and five-membered heteroaromatic rings containingone, two, or three heteroatoms, which are, independently, N, O, or S;alkylaryl, arylalkyl, a C₄₋₁₄ bicyclic ring; a six-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S,

R¹³ is optionally substituted with one or more substituents eachindependently selected from the group consisting of hydrogen, halogen(F, Cl, Br, I), CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂,C(O)R′, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,cycloalkyl, arylalkoxycarbonyl, carboxyl, haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl; or is optionally substituted with aryl,substituted aryl, heteroaryl, and substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl;

or R¹² and R¹³ together with the nitrogen to which they are attachedform a 3 to 4 membered ring optionally substituted with one or moresubstituents each independently selected from the group consisting ofhydrogen, halogen (F, Cl, Br, I), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, and C₁₋₆ hydroxyalkyl.

In a sixth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

K is a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms which are, independently, N, O, or S; a C₄₋₁₄bicyclic ring, alkylheteroaryl, or alkylaryl;

L is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; a four-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, or a C₄₋₁₄ bicyclic ring,

W is

R¹⁴ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹⁵ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ringor a six-membered bridged or spiro-fused ring containing zero, one, ortwo heteroatoms which are, independently, N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms which are,independently, N, O, or S;

R¹⁵ is optionally substituted with one or more substituents which are,independently, halogen (F, Cl, Br, I), SF₅, CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonyl alkyl, C₁₋₆alkyl, cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or is optionally substitutedwith aryl, substituted aryl, heteroaryl, or substituted heteroaryl,where substituents on the substituted aryl and substituted heteroarylare selected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl;

or

R¹⁴ and R¹⁵ can come together with the nitrogen to which they areattached to form a 6-10 membered bicyclic or bridged ring or a 3 to 8saturated ring; such bicyclic, bridged and saturated ring moietyoptionally containing one or more additional heteroatoms which are,independently, O, S or N, and optionally being substituted with one ormore substituents each independently selected from the group consistingof halogen (including F, Cl, Br, and I), SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,and C₁₋₆ hydroxyalkyl.

In a seventh embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

M is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S, aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl,

N is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms independently N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; a four-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S; or a C₄₋₁₄ bicyclic ring,

V is

and

R¹⁶ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, such as phenyl, heteroaryl, such as a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms or afive-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S; a six-membered ring ora six-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S; a four-membered ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S; alkylaryl, arylalkyl,alkylheteroaryl, or alkylaryl,

wherein R¹⁶ is optionally substituted with one or more substituentsselected from the group consisting of halogen (including F, Cl, Br, andI), SF₅, CF₃, hydroxy, N(R)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, hydroxyalkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl.

Representative compounds falling within the scope of the inventioninclude the following:

and pharmaceutically acceptable salts or prodrugs thereof.

Additional compounds also include

and pharmaceutically acceptable salts or prodrugs thereof.

Particularly preferred compounds include:

or a pharmaceutically acceptable salt or prodrug thereof.

Also disclosed are pharmaceutical compositions that include one or moreof the compounds of Formulas I-VII, and a pharmaceutically-acceptablecarrier. The carrier can be, for example, an oral composition, aninjectable composition, a transdermal composition, or a nanoparticulatecomposition. The compositions can further include a second antiviralagent, particularly where the agent is active against HBV infection, andmore particularly where the second antiviral agent is active against HBVinfection via a different mechanism than the instantly-describedcompounds.

Representative types of second antiviral agents include polymeraseinhibitors, viral entry inhibitors, viral maturation inhibitors, capsidassembly modulators, IMPDH inhibitors, protease inhibitors, immune-basedtherapeutic agents, reverse transcriptase inhibitors, TLR-agonists, andagents of distinct or unknown mechanism. Combinations of these agentscan be used.

The compounds described herein can be used to prepare medicaments fortreating HBV infection, preventing an HBV infection, or reducing thebiological activity of an infection with HBV. The medicaments canfurther include another anti-HBV agent.

The compounds and compositions can be used in methods for treating ahost infected with HBV, preventing an infection from a HBV, and reducingthe biological activity of an infection with HBV in a host. The methodscan also involve the co-administration of another anti-HBV agent, whichco-administration can be simultaneous or sequential.

These and other aspects of the invention are further explained in thefollowing detailed description.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows a series of electron micrographs of the result ofincubating HBV Cp149 under conditions that would normally form capsids,and where the incubation was accompanied by the addition of a putativeactive agent, where the active agent functions at least in part byinhibiting capsid formation. Where the incubation was permed usingvehicle alone, the electron micrographs show the capsids in the form offully-formed hollow spheres. When incubated with GLS4, the capsidsformed misassembled hollow spheres, and with Compound 7a, the capsidsformed incomplete hollow spheres, in a relatively low abundance.

FIG. 2 shows a series of electron micrographs of HBV Cp149 capsidstreated with vehicle (showing the capsids in the form of fully-formedhollow spheres), with GLS4, showing that the capsids have formedmisassembled hollow spheres, and with Compound 7a, showing that thecapsids formed incomplete hollow spheres, in a relatively low abundance.

FIG. 3 shows a series of electron micrographs of the capsids shown inFIG. 2, with the first two micrographs repeated, and a third micrographenlarging the portion of the second micrograph to enhance the view ofthe damage to the capsids.

FIG. 4 shows a series of electron micrographs of the capsids shown inFIG. 2, with the first and third micrographs repeated as the first andsecond micrographs. A third micrograph is shown, enlarging the portionof the second micrograph to enhance the view of the damage to thecapsids.

The results show that the compounds effectively disrupted HBV capsidformation.

DETAILED DESCRIPTION

Compounds and compositions useful in treating, preventing, or curing HBVinfection are disclosed. Methods for treating, preventing, or curing HBVinfection are also disclosed.

The compounds described herein show inhibitory activity against HBV incell-based assays. Therefore, the compounds can be used to treat orprevent a HBV in a host, or reduce the biological activity of the virus.The host can be a mammal, and in particular, a human, infected with HBV.The methods involve administering an effective amount of one or more ofthe compounds described herein.

Pharmaceutical formulations including one or more compounds describedherein, in combination with a pharmaceutically acceptable carrier orexcipient, are also disclosed. In one embodiment, the formulationsinclude at least one compound described herein and at least one furthertherapeutic agent.

The present invention will be better understood with reference to thefollowing definitions:

I. Definitions

The term “independently” is used herein to indicate that the variable,which is independently applied, varies independently from application toapplication. Thus, in a compound such as R″XYR″, wherein R″ is“independently carbon or nitrogen,” both R″ can be carbon, both R″ canbe nitrogen, or one R″ can be carbon and the other R″ nitrogen.

As used herein, the term “enantiomerically pure” refers to a compoundcomposition that comprises at least approximately 95%, and, preferably,approximately 97%, 98%, 99% or 100% of a single enantiomer of thatcompound.

As used herein, the term “substantially free of” or “substantially inthe absence of” refers to a compound composition that includes at least85 to 90% by weight, preferably 95% to 98% by weight, and, even morepreferably, 99% to 100% by weight, of the designated enantiomer of thatcompound. In a preferred embodiment, the compounds described herein aresubstantially free of enantiomers.

Similarly, the term “isolated” refers to a compound composition thatincludes at least 85 to 90% by weight, preferably 95% to 98% by weight,and, even more preferably, 99% to 100% by weight, of the compound, theremainder comprising other chemical species or enantiomers.

The term “alkyl,” as used herein, unless otherwise specified, refers toa saturated straight, branched, or cyclic, primary, secondary, ortertiary hydrocarbons, including both substituted and unsubstitutedalkyl groups. The alkyl group can be optionally substituted with anymoiety that does not otherwise interfere with the reaction or thatprovides an improvement in the process, including but not limited to butlimited to halo, haloalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy,amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, aryloxy, nitro, cyano, sulfonic acid, thiol, imine, sulfonyl,sulfanyl, sulfinyl, sulfamonyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid,phosphonate, either unprotected, or protected as necessary, as known tothose skilled in the art, for example, as taught in Greene, et al.,Protective Groups in Organic Synthesis, John Wiley and Sons, SecondEdition, 1991, hereby incorporated by reference. Specifically includedare CF₃ and CH₂CF₃.

In the text, whenever the term C(alkyl range) is used, the termindependently includes each member of that class as if specifically andseparately set out. The term “alkyl” includes C₁₋₂₂ alkyl moieties, andthe term “lower alkyl” includes C₁₋₆ alkyl moieties. It is understood tothose of ordinary skill in the art that the relevant alkyl radical isnamed by replacing the suffix “-ane” with the suffix “-yl”.

As used herein, a “bridged alkyl” refers to a bicyclo- or tricycloalkane, for example, a 2:1:1 bicyclohexane.

As used herein, a “spiro alkyl” refers to two rings that are attached ata single (quaternary) carbon atom.

The term “alkenyl” refers to an unsaturated, hydrocarbon radical, linearor branched, in so much as it contains one or more double bonds. Thealkenyl group disclosed herein can be optionally substituted with anymoiety that does not adversely affect the reaction process, includingbut not limited to but not limited to those described for substituentson alkyl moieties. Non-limiting examples of alkenyl groups includeethylene, methylethylene, isopropylidene, 1,2-ethane-diyl,1,1-ethane-diyl, 1,3-propane-diyl, 1,2-propane-diyl, 1,3-butane-diyl,and 1,4-butane-diyl.

The term “alkynyl” refers to an unsaturated, acyclic hydrocarbonradical, linear or branched, in so much as it contains one or moretriple bonds. The alkynyl group can be optionally substituted with anymoiety that does not adversely affect the reaction process, includingbut not limited to those described above for alkyl moieties.Non-limiting examples of suitable alkynyl groups include ethynyl,propynyl, hydroxypropynyl, butyn-1-yl, butyn-2-yl, pentyn-1-yl,pentyn-2-yl, 4-methoxypentyn-2-yl, 3-methylbutyn-1-yl, hexyn-1-yl,hexyn-2-yl, and hexyn-3-yl, 3,3-dimethylbutyn-1-yl radicals.

The term “alkylamino” or “arylamino” refers to an amino group that hasone or two alkyl or aryl substituents, respectively.

The term “protected” as used herein and unless otherwise defined refersto a group that is added to an oxygen, nitrogen, or phosphorus atom toprevent its further reaction or for other purposes. A wide variety ofoxygen and nitrogen protecting groups are known to those skilled in theart of organic synthesis, and are described, for example, in Greene etal., Protective Groups in Organic Synthesis, supra.

The term “aryl”, alone or in combination, means a carbocyclic aromaticsystem containing one, two or three rings wherein such rings can beattached together in a pendent manner or can be fused. Non-limitingexamples of aryl include phenyl, biphenyl, or naphthyl, or otheraromatic groups that remain after the removal of a hydrogen from anaromatic ring. The term aryl includes both substituted and unsubstitutedmoieties. The aryl group can be optionally substituted with any moietythat does not adversely affect the process, including but not limited tobut not limited to those described above for alkyl moieties.Non-limiting examples of substituted aryl include heteroarylamino,N-aryl-N-alkylamino, N-heteroarylamino-N-alkylamino, heteroaralkoxy,arylamino, aralkylamino, arylthio, monoarylamidosulfonyl,arylsulfonamido, diarylamidosulfonyl, monoaryl amidosulfonyl,arylsulfinyl, arylsulfonyl, heteroarylthio, heteroarylsulfinyl,heteroarylsulfonyl, aroyl, heteroaroyl, aralkanoyl, heteroaralkanoyl,hydroxyaralkyl, hydoxyheteroaralkyl, haloalkoxyalkyl, aryl, aralkyl,aryloxy, aralkoxy, aryloxyalkyl, saturated heterocyclyl, partiallysaturated heterocyclyl, heteroaryl, heteroaryloxy, heteroaryloxyalkyl,arylalkyl, heteroarylalkyl, arylalkenyl, and heteroarylalkenyl,carboaralkoxy.

The terms “alkaryl” or “alkylaryl” refer to an alkyl group with an arylsubstituent. The terms “aralkyl” or “arylalkyl” refer to an aryl groupwith an alkyl substituent.

The term “halo,” as used herein, includes chloro, bromo, iodo andfluoro.

The term “acyl” refers to a carboxylic acid ester in which thenon-carbonyl moiety of the ester group is selected from the groupconsisting of straight, branched, or cyclic alkyl or lower alkyl,alkoxyalkyl, including, but not limited to methoxymethyl, aralkyl,including, but not limited to, benzyl, aryloxyalkyl, such asphenoxymethyl, aryl, including, but not limited to, phenyl, optionallysubstituted with halogen (F, Cl, Br, or I), alkyl (including but notlimited to C₁, C₂, C₃, and C₄) or alkoxy (including but not limited toC₁, C₂, C₃, and C₄), sulfonate esters such as alkyl or aralkylsulphonyl, including but not limited to methanesulfonyl, the mono, di ortriphosphate ester, trityl or monomethoxytrityl, substituted benzyl,trialkylsilyl (e.g., dimethyl-t-butylsilyl) and diphenylmethylsilyl.Aryl groups in the esters optimally comprise a phenyl group. The term“lower acyl” refers to an acyl group in which the non-carbonyl moiety islower alkyl.

The terms “alkoxy” and “alkoxyalkyl” embrace linear or branchedoxy-containing radicals having alkyl moieties, such as methoxy radical.The term “alkoxyalkyl” also embraces alkyl radicals having one or morealkoxy radicals attached to the alkyl radical, that is, to formmonoalkoxyalkyl and dialkoxyalkyl radicals. The “alkoxy” radicals can befurther substituted with one or more halo atoms, such as fluoro, chloroor bromo, to provide “haloalkoxy” radicals. Examples of such radicalsinclude fluoromethoxy, chloromethoxy, trifluoromethoxy, difluoromethoxy,trifluoroethoxy, fluoroethoxy, tetrafluoroethoxy, pentafluoroethoxy, andfluoropropoxy.

The term “alkylamino” denotes “monoalkylamino” and “dialkylamino”containing one or two alkyl radicals, respectively, attached to an aminoradical. The terms arylamino denotes “monoarylamino” and “diarylamino”containing one or two aryl radicals, respectively, attached to an aminoradical. The term “aralkylamino”, embraces aralkyl radicals attached toan amino radical. The term aralkylamino denotes “monoaralkylamino” and“diaralkylamino” containing one or two aralkyl radicals, respectively,attached to an amino radical. The term aralkylamino further denotes“monoaralkyl monoalkylamino” containing one aralkyl radical and onealkyl radical attached to an amino radical.

The term “heteroatom,” as used herein, refers to oxygen, sulfur,nitrogen and phosphorus.

The terms “heteroaryl” or “heteroaromatic,” as used herein, refer to anaromatic that includes at least one sulfur, oxygen, nitrogen orphosphorus in the aromatic ring.

The term “heterocyclic,” “heterocyclyl,” and cycloheteroalkyl refer to anonaromatic cyclic group wherein there is at least one heteroatom, suchas oxygen, sulfur, nitrogen, or phosphorus in the ring.

Nonlimiting examples of heteroaryl and heterocyclic groups includefuryl, furanyl, pyridyl, pyrimidyl, thienyl, isothiazolyl, imidazolyl,tetrazolyl, pyrazinyl, benzofuranyl, benzothiophenyl, quinolyl,isoquinolyl, benzothienyl, isobenzofuryl, pyrazolyl, indolyl,isoindolyl, benzimidazolyl, purinyl, carbazolyl, oxazolyl, thiazolyl,isothiazolyl, 1,2,4-thiadiazolyl, isooxazolyl, pyrrolyl, quinazolinyl,cinnolinyl, phthalazinyl, xanthinyl, hypoxanthinyl, thiophene, furan,pyrrole, isopyrrole, pyrazole, imidazole, 1,2,3-triazole,1,2,4-triazole, oxazole, isoxazole, thiazole, isothiazole, pyrimidine orpyridazine, and pteridinyl, aziridines, thiazole, isothiazole,1,2,3-oxadiazole, thiazine, pyridine, pyrazine, piperazine, pyrrolidine,oxaziranes, phenazine, phenothiazine, morpholinyl, pyrazolyl,pyridazinyl, pyrazinyl, quinoxalinyl, xanthinyl, hypoxanthinyl,pteridinyl, 5-azacytidinyl, 5-azauracilyl, triazolopyridinyl,imidazolopyridinyl, pyrrolopyrimidinyl, pyrazolopyrimidinyl, adenine,N⁶-alkylpurines, N⁶-benzylpurine, N⁶-halopurine, N⁶-vinypurine,N⁶-acetylenic purine, N⁶-acyl purine, N⁶-hydroxyalkyl purine,N⁶-thioalkyl purine, thymine, cytosine, 6-azapyrimidine,2-mercaptopyrmidine, uracil, N⁵-alkylpyrimidines, N⁵-benzylpyrimidines,N⁵-halopyrimidines, N⁵-vinylpyrimidine, N⁵-acetylenic pyrimidine,N⁵-acyl pyrimidine, N⁵-hydroxyalkyl purine, and N⁶-thioalkyl purine, andisoxazolyl. The heteroaromatic group can be optionally substituted asdescribed above for aryl. The heterocyclic or heteroaromatic group canbe optionally substituted with one or more substituents selected fromthe group consisting of halogen, haloalkyl, alkyl, alkoxy, hydroxy,carboxyl derivatives, amido, amino, alkylamino, and dialkylamino. Theheteroaromatic can be partially or totally hydrogenated as desired. As anonlimiting example, dihydropyridine can be used in place of pyridine.Functional oxygen and nitrogen groups on the heterocyclic or heteroarylgroup can be protected as necessary or desired. Suitable protectinggroups are well known to those skilled in the art, and includetrimethylsilyl, dimethylhexylsilyl, t-butyldimethylsilyl, andt-butyldiphenylsilyl, trityl or substituted trityl, alkyl groups, acylgroups such as acetyl and propionyl, methanesulfonyl, andp-toluenelsulfonyl. The heterocyclic or heteroaromatic group can besubstituted with any moiety that does not adversely affect the reaction,including but not limited to but not limited to those described abovefor aryl.

The term “host,” as used herein, refers to a unicellular ormulticellular organism in which the virus can replicate, including butnot limited to cell lines and animals, and, preferably, humans.Alternatively, the host can be carrying a part of the viral genome,whose replication or function can be altered by the compounds of thepresent invention. The term host specifically refers to infected cells,cells transfected with all or part of the viral genome and animals, inparticular, primates (including but not limited to chimpanzees) andhumans. In most animal applications of the present invention, the hostis a human being. Veterinary applications, in certain indications,however, are clearly contemplated by the present invention (such as foruse in treating chimpanzees).

The term “peptide” refers to a natural or synthetic compound containingtwo to one hundred amino acids linked by the carboxyl group of one aminoacid to the amino group of another.

The term “pharmaceutically acceptable salt or prodrug” is usedthroughout the specification to describe any pharmaceutically acceptableform (such as an ester) compound which, upon administration to apatient, provides the compound. Pharmaceutically-acceptable saltsinclude those derived from pharmaceutically acceptable inorganic ororganic bases and acids. Suitable salts include those derived fromalkali metals such as potassium and sodium, alkaline earth metals suchas calcium and magnesium, among numerous other acids well known in thepharmaceutical art.

The term “pharmaceutically acceptable salt or prodrug” is usedthroughout the specification to describe any pharmaceutically acceptableform (such as an ester) compound which, upon administration to apatient, provides the compound. Pharmaceutically acceptable saltsinclude those derived from pharmaceutically acceptable inorganic ororganic bases and acids. Suitable salts include those derived fromalkali metals such as potassium and sodium, alkaline earth metals suchas calcium and magnesium, among numerous other acids well known in thepharmaceutical art. Pharmaceutically acceptable prodrugs refer to acompound that is metabolized, for example hydrolyzed or oxidized, in thehost to form the compound of the present invention. Typical examples ofprodrugs include compounds that have biologically labile protectinggroups on functional moieties of the active compound. Prodrugs includecompounds that can be oxidized, reduced, aminated, deaminated,hydroxylated, dehydroxylated, hydrolyzed, dehydrolyzed, alkylated,dealkylated, acylated, deacylated, phosphorylated, or dephosphorylatedto produce the active compound. The prodrug forms of the compounds ofthis invention can possess antiviral activity, can be metabolized toform a compound that exhibits such activity, or both.

II. Active Compounds

The Hepatitis B virus (HBV) is an enveloped, partially double-strandedDNA (dsDNA) virus of the Hepadnavirus family (Hepadnaviridae). Itsgenome contains 4 overlapping reading frames: the precore/core gene; thepolymerase gene; the L, M, and S genes, which encode for the 3 envelopeproteins; and the X gene.

Upon infection, the partially double-stranded DNA genome (the relaxedcircular DNA; rcDNA) is converted to a covalently closed circular DNA(cccDNA) in the nucleus of the host cell, and the viral mRNAs aretranscribed. Once encapsidated, the pregenomic RNA (pgRNA), which alsocodes for core protein and Pol, serves as the template for reversetranscription, which regenerates the partially dsDNA genome (rcDNA) inthe nucleocapsid.

Following hepatitis B infections, cccDNA can remain following clinicaltreatment in liver cells, and can reactivate. The relative quantity ofcccDNA present is an indicator for HBV treatment (Bourne, et al.,(January 2007). “Quantitative analysis of HBV cccDNA from clinicalspecimens: correlation with clinical and virological response duringantiviral therapy”. Journal of Viral Hepatitis 14 (1): 56-63).

A capsid is the protein shell of a virus, and includes oligomericstructural subunits made of proteins called protomers. The observable3-dimensional morphological subunits, which may or may not correspond toindividual proteins, are called capsomeres. The capsid encloses thegenetic material of the virus.

In vivo, HBV capsids assemble around an RNA-reverse transcriptasecomplex. Assembly of the capsid is required for reverse transcription ofthe RNA pregenome to the mature DNA form. In HBV, the dominant form ofcapsid is composed of 120 copies of the capsid protein dimer. Evenmodest mutations of the capsid protein can have dramatic effects on theviability of progeny virus.

Most of the compounds described herein are active as capsid inhibitors.Inhibiting capsid assembly can reduce cccDNA, the main reservoir forHBV, and can also decrease the levels of HBV DNA, HBeAg and HBsAg.

In one embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

A is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl;

B is a six or seven-membered ring or a six or seven-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S, a five-membered ring containing zero, one, ortwo heteroatoms, which are, independently, N, O, or S; a four-memberedring containing zero, one, or two heteroatoms, which are, independently,N, O, or S, or a C₄₋₁₄ bicyclic ring,

When R¹ and R^(1′) are attached to a carbon, they are, independently,hydrogen, halogen (including F, Cl, Br, and I), CF₃, SF₅, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₂₋₆ alkenyl, cyano,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl,C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl;

When R¹ and R^(1′) are attached to a nitrogen, they are, independently,hydrogen, C₂₋₆ alkoxy, C₃₋₆ alkoxyalkyl, C₂₋₆ alkenyl, alkoxycarbonyl,carbonylalkyl, carbonyl aryl, C₁₋₆ alkyl, heterocyclylalkyl, C₂₋₆hydroxyalkyl, or S(O)₂R′;

Each R′ is independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl,alkylaryl, or arylalkyl, or if two R′ reside on the same nitrogen atom,they can come together to form a C₃₋₆ ring optionally containing a N, O,or S heteroatom;

The R′ groups can optionally be substituted with one or moresubstituents, which substituents are, independently, halo, C₁₋₆haloalkyl, C₁₋₆ hydroxyalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy,amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, alkoxyalkyl, aryloxy, nitro, cyano, sulfonic acid, thiol, imine,sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid, orphosphonate, either unprotected, or protected as necessary, as known tothose skilled in the art, for example, as taught in Greene, et al.,Protective Groups in Organic Synthesis, John Wiley and Sons, SecondEdition, 1991, hereby incorporated by reference;

u and v are independently 0, 1, 2, 3, 4 or 5;

X is

R³ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R² is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, such as phenyl, heteroaryl, including six-memberedheteroaromatic rings containing one, two, or three nitrogen atoms andfive-membered heteroaromatic rings containing one, two, or threeheteroatoms, which, independently, are N, O, or S, alkylaryl, arylalkyl,a six-membered ring or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, a seven-membered bridged or spiro-fused ring containing zero, one,or two heteroatoms which are, independently, N, O, or S, a five-memberedring containing zero, one, or two heteroatoms which are, independently,N, O, or S; a four-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; cycloalkyl,alkylheteroaryl, or alkylaryl;

R² is optionally substituted with one or more substituents, which each,independently, are halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or issubstituted with aryl, substituted aryl, heteroaryl, or substitutedheteroaryl, where substituents on the substituted aryl and substitutedheteroaryl are selected from the group consisting of halogen, SF₅, CF₃,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano,azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, and C₁₋₆ alkyl.

R² and R³ can come together with the nitrogen to which they are attachedform a 6-10 membered bicyclic or bridged ring, a 3 to 8 saturated ring,or a 5 membered unsaturated ring; such bicyclic, bridged, saturated andunsaturated rings optionally containing one or more additionalheteroatoms, where each is, independently, O, S or N, and optionallybeing substituted with one or more substituents, wherein each,independently, is halogen (including F, Cl, Br, I), CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl.

In a second embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

C is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylaryl, or alkylheteroaryl;

D is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S, ora C₄₋₁₄ bicyclic ring,

Y is

R⁴ is H or C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl; inone embodiment, R⁴ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl,

where, in one embodiment, if C is phenyl, D is not phenyl or a5-membered ring heteroaryl, and in another embodiment, if C is phenyland D is phenyl or a 5-membered ring heteroaryl, then R⁵ is notalkylaryl, alkenyl, or a six-membered bridged ring;

R⁵ is alkylaryl, arylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, such asphenyl, heteroaryl, including six-membered heteroaromatic ringscontaining one, two, or three nitrogen atoms and five-memberedheteroaromatic rings containing one, two, or three heteroatoms, which,independently, are N, O, or S; and a six-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S; in one embodiment, R⁵ is alkylaryl, arylalkyl, phenyl, afive or six-membered heteroaryl, or a six-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S;

R⁵ is optionally substituted with one or more substituents, each ofwhich is, independently, halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclyl alkyl, or C₁₋₆hydroxyalkyl; or is substituted with aryl, substituted aryl, heteroaryl,or substituted heteroaryl, where substituents on the substituted aryland substituted heteroaryl are selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl;

or when Y is

R⁴ and R⁵ together with the nitrogen to which they are attached form a 3to 4 membered ring optionally substituted with one or more substituents,each of which is, independently, halogen (including F, Cl, Br, I), CF₃,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl,C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In one embodiment of the compounds of Formula II, D is

where R⁶ is H, Cl, F or Br, and R⁷ is H, methyl, F or Cl.

In one aspect of this embodiment, when Y is

R⁵ is not

In another aspect of this embodiment, when R⁴ is ethyl, then R⁵ is not

In one embodiment of the compounds of Formula II, C is a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S; a C₄₋₁₄ bicyclic ring,alkylaryl, or alkylheteroaryl.

In one embodiment of the compounds of Formula II, D is a C₄₋₁₄ bicyclicring.

In another embodiment of the compounds of Formula II, R⁵ is arylalkyl,C₂₋₆ alkynyl, aryl, such as phenyl, heteroaryl, including six-memberedheteroaromatic rings containing one, two, or three nitrogen atoms andfive-membered heteroaromatic rings containing one, two, or threeheteroatoms, which, independently, are N, O, or S; and a six-memberedspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S.

In a third embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

E is a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms, where each is, independently, N, O, or S; a C₄₋₁₄bicyclic ring, alkylheteroaryl, or alkylaryl;

F is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S, or a C₄₋₁₄ bicyclicring,

Z is

R⁸ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R⁹ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ringor a six-membered bridged or spiro-fused ring containing zero, one, ortwo heteroatoms, which are independently N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms, which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S, or a three membered ring;

R⁹ is optionally substituted with one or more substituents, each ofwhich is independently halogen (including F, Cl, Br, and I), CF₃, SF₅,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, C₁₋₆hydroxyalkyl; or is substituted with aryl, substituted aryl, heteroaryl,or substituted heteroaryl, where substituents on the substituted aryland substituted heteroaryl are selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl.

R⁸ and R⁹ can come together with the nitrogen to which they are attachedform a 6-10 membered bicyclic or bridged ring or a 3 to 8 saturatedring; such bicyclic, bridged and saturated ring moiety optionallycontaining one or more additional heteroatoms which, independently, areO, S or N and optionally being substituted with one or moresubstituents, each, independently, is halogen (including F, Cl, Br, andI), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In a fourth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

G is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S; aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl;

H is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a six-membered non-aromatic ring optionallycontaining one, two, or three heteroatoms, which are, independently, N,O, or S; or a C₄₋₁₄ bicyclic ring;

When R¹ and R^(1′) are attached to a carbon they are, independently,hydrogen, halogen (including F, Cl, Br, and I), CF₃, SF₅, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, C₂₋₆ alkynyl,C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl;

When R¹ and R^(1′) are attached to a nitrogen they are, independently,hydrogen, C₁₋₆ alkoxy, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, carbonylalkyl,carbonyl aryl, C₁₋₆ alkyl, C₂₋₆ alkynyl, C₂₋₆ alkenyl,heterocyclylalkyl, C₁₋₆ hydroxyalkyl, or S(O)₂R′;

Each R′ is independently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy,C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl,alkylaryl, or arylalkyl, or if two R′ reside on the same nitrogen atomthey can come together to form a C₃₋₆ alkyl ring optionally containing aN, O, or S; wherein the R′ groups can be substituted with one or moresubstituents as defined above, for example, C₁₋₆ hydroxyalkyl,aminoalkyl, and alkoxyalkyl;

u and v are independently 0, 1, 2, 3, 4 or 5;

W is

R¹⁰ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹¹ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, heteroaryl, alkylaryl, arylalkyl, phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, asix-membered ring or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S, a seven-membered bridged or spiro-fused ring containing zero,one, or two heteroatoms, which are, independently, N, O, or S, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S; a four-membered ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S; a three membered ring,alkylheteroaryl, or alkylaryl;

wherein R¹¹ is optionally substituted with one or more substituentsselected from the group consisting of halogen (including F, Cl, Br, andI), SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy,C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, C₁₋₆ hydroxyalkyl, aryl,substituted aryl, heteroaryl, and substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl; or

R¹⁰ and R¹¹ can come together with the nitrogen to which they areattached form a 6-10 membered bicyclic or bridged ring or a 3 to 8saturated ring; such bicyclic, bridged or saturated ring moietyoptionally containing one or more additional heteroatoms, which areeach, independently, O, S or N, and optionally substituted with one ormore substituents, each of which is, independently, halogen (includingF, Cl, Br, and I), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂,C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl.

In a fifth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

I is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms, which are, independently, N, O, or S, aC₄₋₁₄ bicyclic ring; alkylheteroaryl, or alkylaryl;

J is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; or a four-memberedring containing zero, one, or two heteroatoms which are, independently,N, O, or S,

W is

R¹² is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹³ is C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, including phenyl, heteroaryl,including six-membered heteroaromatic rings containing one, two, orthree nitrogen atoms and five-membered heteroaromatic rings containingone, two, or three heteroatoms, which are, independently, N, O, or S;alkylaryl, arylalkyl, a C₄₋₁₄ bicyclic ring; a six-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S,

R¹³ is optionally substituted with one or more substituents eachindependently selected from the group consisting of hydrogen, halogen(F, Cl, Br, I), CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂,C(O)R′, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆, alkyl,cycloalkyl, arylalkoxycarbonyl, carboxyl, haloalkyl, heterocyclylalkyl,and C₁₋₆ hydroxyalkyl; or is optionally substituted with aryl,substituted aryl, heteroaryl, or substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl;

or R¹² and R¹³ together with the nitrogen to which they are attachedform a 3 to 4 membered ring optionally substituted with one or moresubstituents each independently selected from the group consisting ofhydrogen, halogen (F, Cl, Br, I), CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, haloalkyl, heterocyclylalkyl, and C₁₋₆ hydroxyalkyl.

In a sixth embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

K is a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms which are, independently, N, O, or S; a C₄₋₁₄bicyclic ring, alkylheteroaryl, or alkylaryl;

L is a five-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; a four-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, or a C₄₋₁₄ bicyclic ring,

W is

R¹⁴ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl,

R¹⁵ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ringor a six-membered bridged or spiro-fused ring containing zero, one, ortwo heteroatoms which are, independently, N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms which are,independently, N, O, or S;

R¹⁵ is optionally substituted with one or more substituents which are,independently, halogen (F, Cl, Br, I), SF₅, CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, and C₁₋₆ hydroxyalkyl; or is substituted with aryl,substituted aryl, heteroaryl, or substituted heteroaryl, wheresubstituents on the substituted aryl and substituted heteroaryl areselected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl, or

R¹⁴ and R¹⁵ can come together with the nitrogen to which they areattached form a 6-10 membered bicyclic or bridged ring or a 3 to 8saturated ring; such bicyclic, bridged and saturated ring moietyoptionally containing one or more additional heteroatoms which are,independently, O, S or N, and optionally being substituted with one ormore substituents each independently selected from the group consistingof halogen (including F, Cl, Br, and I), CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, and C₁₋₆hydroxyalkyl.

In a seventh embodiment, the compounds have the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein

R¹ and R^(1′) are as defined with respect to Formula I,

u and v are independently 0, 1, 2, 3, 4 or 5;

M is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms which are, independently, N, O, or S, aC₄₋₁₄ bicyclic ring, alkylheteroaryl, or alkylaryl,

N is phenyl, a six-membered heteroaromatic ring containing one, two, orthree nitrogen atoms, a five-membered heteroaromatic ring containingone, two, or three heteroatoms independently selected from N, O, and S,a six or seven-membered ring or a six or seven-membered bridged orspiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, or S, a five-membered ring containing zero, one, ortwo heteroatoms which are, independently, N, O, or S; a four-memberedring containing zero, one, or two heteroatoms which are, independently,N, O, or S; or a C₄₋₁₄ bicyclic ring,

V is

and

R¹⁶ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, such as phenyl, heteroaryl, such as a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms or afive-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S; a six-membered ring ora six-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S; a four-membered ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S; alkylaryl, arylalkyl,alkylheteroaryl, or alkylaryl, wherein R¹⁶ is optionally substitutedwith one or more substituents selected from the group consisting ofhalogen (including F, Cl, Br, and I), SF₅, CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl,C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, C₁₋₆ hydroxyalkyl, aryl, substituted aryl,heteroaryl, and substituted heteroaryl, where substituents on thesubstituted aryl and substituted heteroaryl are selected from the groupconsisting of halogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl.

Representative compounds falling within the scope of the inventioninclude the following:

and pharmaceutically acceptable salts or prodrugs thereof.

Representative compounds also include:

and pharmaceutically acceptable salts and prodrugs thereof.

Particularly preferred compounds include:

or a pharmaceutically acceptable salt or prodrug thereof.

A particularly preferred compound has the formula:

or a pharmaceutically acceptable salt thereof.

III Stereoisomerism and Polymorphism

The compounds described herein can have asymmetric centers and occur asracemates, racemic mixtures, individual diastereomers or enantiomers,with all isomeric forms being included in the present invention.Compounds of the present invention having a chiral center can exist inand be isolated in optically active and racemic forms. Some compoundscan exhibit polymorphism. The present invention encompasses racemic,optically-active, polymorphic, or stereoisomeric forms, or mixturesthereof, of a compound of the invention, which possess the usefulproperties described herein. The optically active forms can be preparedby, for example, resolution of the racemic form by recrystallizationtechniques, by synthesis from optically-active starting materials, bychiral synthesis, or by chromatographic separation using a chiralstationary phase or by enzymatic resolution. One can either purify therespective compound, then derivatize the compound to form the compoundsdescribed herein, or purify the compound themselves.

Optically active forms of the compounds can be prepared using any methodknown in the art, including but not limited to by resolution of theracemic form by recrystallization techniques, by synthesis fromoptically-active starting materials, by chiral synthesis, or bychromatographic separation using a chiral stationary phase.

Examples of methods to obtain optically active materials include atleast the following.

i) physical separation of crystals: a technique whereby macroscopiccrystals of the individual enantiomers are manually separated. Thistechnique can be used if crystals of the separate enantiomers exist,i.e., the material is a conglomerate, and the crystals are visuallydistinct;

ii) simultaneous crystallization: a technique whereby the individualenantiomers are separately crystallized from a solution of the racemate,possible only if the latter is a conglomerate in the solid state;

iii) enzymatic resolutions: a technique whereby partial or completeseparation of a racemate by virtue of differing rates of reaction forthe enantiomers with an enzyme;

iv) enzymatic asymmetric synthesis: a synthetic technique whereby atleast one step of the synthesis uses an enzymatic reaction to obtain anenantiomerically pure or enriched synthetic precursor of the desiredenantiomer;

v) chemical asymmetric synthesis: a synthetic technique whereby thedesired enantiomer is synthesized from an achiral precursor underconditions that produce asymmetry (i.e., chirality) in the product,which can be achieved using chiral catalysts or chiral auxiliaries;

vi) diastereomer separations: a technique whereby a racemic compound isreacted with an enantiomerically pure reagent (the chiral auxiliary)that converts the individual enantiomers to diastereomers. The resultingdiastereomers are then separated by chromatography or crystallization byvirtue of their now more distinct structural differences and the chiralauxiliary later removed to obtain the desired enantiomer;

vii) first- and second-order asymmetric transformations: a techniquewhereby diastereomers from the racemate equilibrate to yield apreponderance in solution of the diastereomer from the desiredenantiomer or where preferential crystallization of the diastereomerfrom the desired enantiomer perturbs the equilibrium such thateventually in principle all the material is converted to the crystallinediastereomer from the desired enantiomer. The desired enantiomer is thenreleased from the diastereomer;

viii) kinetic resolutions: this technique refers to the achievement ofpartial or complete resolution of a racemate (or of a further resolutionof a partially resolved compound) by virtue of unequal reaction rates ofthe enantiomers with a chiral, non-racemic reagent or catalyst underkinetic conditions;

ix) enantiospecific synthesis from non-racemic precursors: a synthetictechnique whereby the desired enantiomer is obtained from non-chiralstarting materials and where the stereochemical integrity is not or isonly minimally compromised over the course of the synthesis;

x) chiral liquid chromatography: a technique whereby the enantiomers ofa racemate are separated in a liquid mobile phase by virtue of theirdiffering interactions with a stationary phase (including but notlimited to via chiral HPLC). The stationary phase can be made of chiralmaterial or the mobile phase can contain an additional chiral materialto provoke the differing interactions;

xi) chiral gas chromatography: a technique whereby the racemate isvolatilized and enantiomers are separated by virtue of their differinginteractions in the gaseous mobile phase with a column containing afixed non-racemic chiral adsorbent phase;

xii) extraction with chiral solvents: a technique whereby theenantiomers are separated by virtue of preferential dissolution of oneenantiomer into a particular chiral solvent;

xiii) transport across chiral membranes: a technique whereby a racemateis placed in contact with a thin membrane barrier. The barrier typicallyseparates two miscible fluids, one containing the racemate, and adriving force such as concentration or pressure differential causespreferential transport across the membrane barrier. Separation occurs asa result of the non-racemic chiral nature of the membrane that allowsonly one enantiomer of the racemate to pass through.

Chiral chromatography, including but not limited to simulated moving bedchromatography, is used in one embodiment. A wide variety of chiralstationary phases are commercially available.

IV. Salt or Prodrug Formulations

In cases where compounds are sufficiently basic or acidic to form stablenontoxic acid or base salts, administration of the compound as apharmaceutically acceptable salt may be appropriate. Examples ofpharmaceutically acceptable salts are organic acid, which form aphysiological acceptable anion, for example, tosylate, methanesulfonate,acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate,α-ketoglutarate and α-glycerophosphate. Suitable inorganic salts canalso be formed, including but not limited to, sulfate, nitrate,bicarbonate and carbonate salts. For certain transdermal applications,it can be preferred to use fatty acid salts of the compounds describedherein. The fatty acid salts can help penetrate the stratum corneum.Examples of suitable salts include salts of the compounds with stearicacid, oleic acid, lineoleic acid, palmitic acid, caprylic acid, andcapric acid.

Pharmaceutically acceptable salts can be obtained using standardprocedures well known in the art, for example by reacting a sufficientlybasic compound such as an amine with a suitable acid, affording aphysiologically acceptable anion. In those cases where a compoundincludes multiple amine groups, the salts can be formed with any numberof the amine groups. Alkali metal (e.g., sodium, potassium or lithium)or alkaline earth metal (e.g., calcium) salts of carboxylic acids canalso be made.

A prodrug is a pharmacological substance that is administered in aninactive (or significantly less active) form and subsequentlymetabolized in vivo to an active metabolite. Getting more drug to thedesired target at a lower dose is often the rationale behind the use ofa prodrug and is generally attributed to better absorption,distribution, metabolism, and/or excretion (ADME) properties. Prodrugsare usually designed to improve oral bioavailability, with poorabsorption from the gastrointestinal tract usually being the limitingfactor. Additionally, the use of a prodrug strategy can increase theselectivity of the drug for its intended target thus reducing thepotential for off target effects.

V. Isotopes

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulae and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. In other embodimentsare examples of isotopes that are incorporated into the presentcompounds including isotopes of hydrogen, carbon, nitrogen, oxygen,fluorine and chlorine, such as, for example, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O,¹⁷O, ³⁵S, ¹⁸F, ³⁶Cl, respectively. Certain isotopically-labeledcompounds described herein, for example those into which radioactiveisotopes such as ²H are incorporated, are useful in drug and/orsubstrate tissue distribution assays. Further, in some embodiments,substitution with isotopes such as deuterium, i.e., ²H, can affordscertain therapeutic advantages resulting from greater metabolicstability, such as, for example, increased in vivo half-life or reduceddosage requirements.

VI. Methods of Treatment

The compounds described herein can be used to prevent, treat or curehepatitis B virus (HBV) infections and West Nile virus infections.

Hosts, including but not limited to humans, suffering from one of thesecancers, or infected with one of these viruses, such as HBV, or a genefragment thereof, can be treated by administering to the patient aneffective amount of the active compound or a pharmaceutically acceptableprodrug or salt thereof in the presence of a pharmaceutically acceptablecarrier or diluent. The active materials can be administered by anyappropriate route, for example, orally, parenterally, intravenously,intradermally, transdermally, subcutaneously, or topically, in liquid orsolid form.

The compounds and compositions described herein can also be used totreat other viral diseases. For example, by curing, controlling, oreliminating HBV, HDV infection can also be suppressed or eliminated.Sheldon et al., “Does treatment of hepatitis B virus (HBV) infectionreduce hepatitis delta virus (HDV) replication in HIV-HBV-HDV-coinfectedpatients?” Antivir Ther. 2008; 13(1):97-102).

Hepatitis delta virus (HDV) has a unique replication process thatrequires coinfection with hepatitis B virus (HBV). While treatment isbelieved to be currently limited to interferon therapy, patientsundergoing successful anti-HBV therapy with the compounds describedherein can indirectly benefit from suppression of HDV replication. Asignificant and sustained reduction in serum HDV RNA can be obtained byreducing the HBV covalently closed circular DNA (cccDNA). cccDNA in HBVis formed by conversion of capsid-associated relaxed circular DNA(rcDNA). [Guo et al., “Characterization of the intracellulardeproteinized relaxed circular DNA of hepatitis B virus: an intermediateof covalently closed circular DNA formation”. J Virol. 81 (22):12472-12484 (November 2007). Accordingly, inhibition of capsid formationusing the compounds described herein can also suppress or eliminate HDVreplication.

Further, there is a subset of HCV patients which have also been earlierinfected with HBV, and the HBV is dormant at the time the HCV is beingtreated. in some of these patients, successful treatment of HCV (forexample, with Harvoni/Sovaldi) can reactivate the dormant HBV infection.Co-administration of the compounds described herein, along with HCVtreatment, can prevent the reactivation of the dormant HBV infection, ortreat the reactivated HBV infection.

VII. Combination of Alternation Therapy

In one embodiment, the compounds of the invention can be employedtogether with at least one other antiviral agent, including, but notlimited to, polymerase inhibitors, anti-HBV nucleosides and theirprodrugs, viral entry inhibitor, viral maturation inhibitor, literaturedescribed capsid assembly modulator, IMPDH inhibitors, proteaseinhibitors, immune-based therapeutic agents, reverse transcriptaseinhibitor, a TLR-agonist, and agents of distinct or unknown mechanism.They can also be used in conjunction with CRISPR/CAS9 approaches usingAAV as the human delivery vector.

For example, when used to treat or prevent HBV infection, the activecompound or its prodrug or pharmaceutically acceptable salt can beadministered in combination or alternation with another anti-HBV agentincluding, but not limited to, those of the formula above. In general,in combination therapy, effective dosages of two or more agents areadministered together, whereas during alternation therapy, an effectivedosage of each agent is administered serially. The dosage will depend onabsorption, inactivation and excretion rates of the drug, as well asother factors known to those of skill in the art. It is to be noted thatdosage values will also vary with the severity of the condition to bealleviated. It is to be further understood that for any particularsubject, specific dosage regimens and schedules should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecompositions.

Nonlimiting examples of antiviral agents that can be used in combinationwith the compounds disclosed herein include those in the tables below.

Hepatitis B Therapies FAMILY/DRUG COMPANY/ NAME MECHANISM STATUS IntronA Immunomodulator Merck, (Interferon Whitehouse alfa-2b) Station, NJApproved Pegasys Immunomodulator Genentech, (Peginterferon South Sanalfa-2a) Francisco, CA Approved Epivir-HBV Inhibits viral DNAGlaxoSmithKline, (Lamivudine) polymerase Philadelphia, PA ApprovedHepsera Inhibits viral DNA Gilead Sciences, (Adefovir polymerase FosterCity, CA Dipivoxil) Approved Baraclude Inhibits viral DNA Bristol-Myers(Entecavir) polymerase Squibb, Princeton, NJ Approved Tyzeka Inhibitsviral DNA Novartis, (Telbivudine) polymerase Switzerland Approved VireadInhibits viral DNA Gilead Sciences, (Tenofovir) polymerase Foster City,CA Approved Clevudine Inhibits viral DNA Bukwang, (L-FMAU) polymeraseSouth Korea Eisai, Japan Approved S. Korea 2006 (Levovir) TenofovirProdrug of Tenofovir Gilead Sciences, alafenamide Foster City, CA (TAF)Phase III CMX157 Prodrug of Tenofovir ContraVir Pharmaceuticals, Edison,NJ Phase II AGX-1009 Prodrug of Tenofovir Agenix, Australia Phase II,China Myrcludex B Entry Inhibitor Hepatera, Russia With Myr-GmbH,Germany Phase II for HBV and HDV ARC520 RNAi gene silencer ArrowheadResearch, Pasadena, CA Phase II/III NVR 3-778 Capsid Inhibitor NoviraTherapeutics, Doylestown, PA Phase IIa Morphothiadine Capsid inhibitorSunshine Lake mesilate (GLS4) Pharma of HEC, China Phase II ISIS-HBVRxAntisense drug ISIS Pharma (w/ GSK), Carlsbad, CA Phase II SB 9200 HBVSmall molecule Spring Bank nucleic acid hybrids Pharma, or “SMNH”Milford, MA Phase II Rep 2139-Ca HBsAg release REPLIC or Inc., inhibitorCanada Phase II Bay 41-4109 Capsid Inhibitor AiCuris, Germany Phase ITKM-HBV RNAi gene silencer Tekmira, Canada Phase I Alinia Small moleculeRomark Labs, (Nitazoxanide) Tampa, FL Preclinical CpAMS HBV Core ProteinAssembly Biosciences, NY, NY Preclinical ALN-HBV RNAi gene silencerAlnylam, Cambridge, MA PreclinicaL CPI-431-32 Cyclophilin inhibitorCiclofilin Pharma, San Diego, CA Preclinical Hepbarna RNAi genesilencing Benitec, Australia preclinical OCB-030 Cyclophilin inhibitorArbutus Biopharma (formerly Tekmira), Canada Preclinical GS-9620 TLR7agonist Gilead Sciences, Foster City, CA′ Phase II RG7795 TLR7 agonistRoche, Switzerland (formerly Phase II ANA773) CYT107 ImmunomodulatorCytheris, Interleukin-7) France Phase I/IIa NCT01641536 Therapeuticvaccine Ichor Medical Systems (w/Janssen), San Diego, CA Phase I TG 1050Immunotherapeutic Transgene, Shanghai Phase I CYT-003 TLR9 agonistArbutus Biopharma (formerly Tekmira), Canada Preclinical ARB 1467TKM-HBV Arbutus Biopharma (formerly Tekmira), Canada PreclinicalARB-1468 TKM-HBV Arbutus Biopharma, (formerly Tekmira), CanadaPreclinical

Additional Anti-HBV Treatments which can be Used in Combination orAlternation

In addition to the compounds described herein, which can function byinhibiting cccDNA, and the combination therapies described above, whichcombine the compounds described herein with approved anti-HBV drugs suchas TAF, approaches like siRNA, shRNA, Talens, Crisper/Cas9, and mir(microRNA) compounds can also be used.

siRNA and shRNA Therapy

siRNA therapy for treating HBV is described, for example, in Chen andMahato, “siRNA Pool Targeting Different Sites of Human Hepatitis BSurface Antigen Efficiently Inhibits HBV Infection;” J Drug Target. 2008February; 16(2): 140-148 and Morrissey et al., “Potent and persistent invivo anti-HBV activity of chemically modified siRNAs,” NatureBiotechnology 23, 1002-1007 (2005).

RNAi is a sequence-specific, post-transcriptional gene silencingmechanism, which is triggered by double-stranded synthetic siRNA orshort hairpin RNA (shRNA) expressed intracellularly from a vector. HBVreplication and expression can be inhibited by administration ofsynthetic siRNAs or endogenously expressed shRNAs. See, for example,Giladi et al., “Small interfering RNA inhibits hepatitis B virusreplication in mice,” Mol Ther. 2003; 8(5):769-76; McCaffrey et al.,“Inhibition of hepatitis B virus in mice by RNA interference,” NatBiotechnol. 2003; 21(6):639-44; and Shlomai and Shaul, “Inhibition ofhepatitis B virus expression and replication by RNA interference,”Hepatology. 2003; 37(4):764-70). HBV gene silencing may depend, forexample, on siRNA dosing and sequences, and targets for gene silencinginclude, for example, the inhibition of virus replication, andsuppression of HBsAg expression.

In one embodiment, a combination of several siRNAs and/or shRNAs areused, targeting two or more of the HBV S, C, P and X genes. In thismanner, multiple targets for inhibition of HBV replication and geneexpression can be accessed.

Once an appropriate target has been identified, for example, the humanhepatitis B virus surface antigen (HBsAg) (Gene Bank Accession#NM_U95551), siRNAs can be designed according to the guide provided byAmbion (http://www.ambion.com/techlib/misc/siRNA_finder.html) andInvitrogen (https://rnaidesigner.invitrogen.com/rnaiexpress/design.do).The sequence specificity of siRNAs can be checked by performing a BLASTsearch (www.ncbi.nlm.nih.gov).

Once siRNA sequences are identified, they can be converted into shRNAs.To express shRNA, control vectors can be constructed, for example, usingpsiSTRIKE™, which is a linearized plasmid and contains a U6 RNApolymerase promoter. These shRNAs contain two complementaryoligonucleotides that can be annealed to form double-stranded DNA forligation into psiSTRIKE™ vector corresponding sites, under a suitablepromoter, such as the U6 promoter, using an appropriate ligase, such asT4 DNA ligase. Plasmids can be purified, for example, using the QIAGEN®Plasmid Mini Kit (QIAGEN, Valencia, Calif.).

Talens/CRISPR

As discussed above, chronic HBV viral infections often persist due tothe presence of long-lived forms of viral DNA in infected cells. Currenttherapies can suppress viral replication, but have little or no effecton long-lived DNA forms, so viral replication resumes as soon as therapyis stopped.

In addition to targeting long-lived DNA forms using the capsidinhibitors described herein, targeted endonucleases, such as homingendonucleases, zinc-finger nucleases (ZFNs), transcriptionactivator-like effector nucleases (TALENs), and the CRISPR (clusteredregularly interspaced short palindromic repeats) system can be used. Theuse of TALENS to target HBV is described, for example, in Weber et al.,“TALENs Targeting HBV: Designer Endonuclease Therapies for ViralInfections,” Molecular Therapy (2013); 21 10, 1819-1820;http://www.nature.com/mt/journal/v21/n10/full/mt2013208a.html

These nucleases function by specifically recognizing and cleavingselected DNA sequences, which results in gene disruption upon impreciseDNA repair. TALENs targeting of the hepatitis B virus (HBV) genome canresult in TALEN-induced mutations in the long-lived HBV covalentlyclosed circular DNA (cccDNA). Mutation and/or disruption of cccDNAprevents viral replication by blocking expression of functional viralproteins.

CRISPR

CRISPR, or clustered regularly interspaced short palindromic repeats, isanother way to mutate HBV DNA, by providing targeted genome editing. Inaddition to the programmable editing tools, such as zinc fingernucleases and transcription activator-like effector nucleases (TALENs)described above, CRISPR (clustered regularly interspaced shortpalindromic repeats)/Cas9 technology also allows for genome editing, andallows for site-specific genomic targeting in HBV.

The type II CRISPR/Cas system is a prokaryotic adaptive immune responsesystem that uses non-coding RNAs to guide the Cas9 nuclease to inducesite-specific DNA cleavage. This DNA damage is repaired by cellular DNArepair mechanisms, either via the non-homologous end joining DNA repairpathway (NHEJ) or the homology directed repair (HDR) pathway.

The CRISPR/Cas9 system provides a simple, RNA-programmable method togenerate gene knockouts (via insertion/deletion) or knockins (via HDR),and allows for site-specific genomic targeting in HBV. The type IICRISPR/Cas system is a prokaryotic adaptive immune response system thatuses non-coding RNAs to guide the Cas9 nuclease to induce site-specificDNA cleavage.

To create gene disruptions, a single guide RNA (sgRNA) is generated todirect the Cas9 nuclease to a specific genomic location. Cas9-induceddouble strand breaks are repaired via the NHEJ DNA repair pathway. Therepair is error prone, and thus insertions and deletions (INDELs) may beintroduced that can disrupt gene function.

Thus, targeting hepatitis B virus cccDNA using a CRISPR/Cas9 nucleasecan efficiently inhibits viral replication.

Mir/MicroRNA

MicroRNAs (miRNAs) are tiny noncoding RNAs that regulate gene expressionprimarily at the post-transcriptional level by binding to mRNAs. miRNAscontribute to a variety of physiological and pathological processes. Anumber of miRNAs have been found to play a pivotal role in the host-HBVinteraction. HBV infection can change the cellular miRNA expressionpatterns, and different stages of HBV associated disease have displayeddistinctive miRNA profiles. The differential expressed miRNAs areinvolved in the progression of HBV-related diseases. For instance, somemiRNAs are involved in liver tumorigenesis and tumor metastasis.Circulating miRNA in serum or plasma can be a very useful biomarker forthe diagnosis and prognosis of HBV-related diseases. In addition,miRNA-based therapy can be used to treat, prevent, or cure HBV-relateddiseases. See, for example, Ying-Feng Wei, “MicroRNAs may solve themystery of chronic hepatitis B virus infection,” World J Gastroenterol.2013 Aug. 14; 19(30): 4867-4876.http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3740416/

In the interaction between virus and host, miRNAs can be divided intocellular miRNAs and viral miRNAs. Cellular miRNAs' expression profileschange at the infected state, and abnormal miRNAs often closely relateto the viral life cycle as well as the host disorder. Viral miRNAs canevolve to regulate both viral and cellular gene expression.

Sometimes, viruses exploit cellular miRNAs to facilitate certain stepsof their life cycle. For example, miR-122 serves an antiviral role inthe HBV life cycle. MiR-122 over-expression inhibits HBV expression,whereas depletion of endogenous miR-122 results in increased HBVproduction in transfected cells. MiR-122 inhibitors cause an increase incellular heme oxygenase-1, which can decrease HBV covalently closedcircular DNA (cccDNA) levels by reducing the stability of the HBV coreprotein. MiR-122 expression in the liver can be significantlydown-regulated in patients with HBV infection compared with healthycontrols. MiR-122 is significantly up-regulated in HBV-infectedpatients, and can inhibit HBV replication in Huh7 and HepG2 cells.Cyclin G1 is a miR-122 target that specifically interacts with p53,resulting in the specific binding of p53 to the HBV enhancer elementsand simultaneous abrogation of the p53-mediated inhibition of HBVtranscription.

HBV is a noncytopathic virus that replicates preferentially in thehepatocytes. cccDNA serves as a template for transcription of all viralRNA that is synthetized after HBV DNA enters the hepatocyte nucleus. TheHBV genome is 3.2 kb in length and contains four overlapping openreading frames. It can transcribe viral pregenomic RNA that reversestranscription to synthesize the viral DNA genome and encode thehepatitis B virus surface antigen (HBsAg), hepatitis B virus coreprotein, viral reverse DNA polymerase (Pol) and X protein.

Hsa-miR-125a-5p interferes with HBV translation and down-regulates theexpression of the HBV surface antigen. Accordingly, cellular miRNAs canalter HBV gene expression by targeting to HBV transcripts.

Cellular miRNAs can affect viral translation and change viralreplication. In addition to the instance of the miR-122 inhibition ofHBV replication, there are other examples where host miRNAs alter HBVreplication. MiR-141 suppresses HBV replication by reducing HBV promoteractivities, by down-regulating peroxisome proliferator-activatedreceptor alpha. DNA hypermethylation may be closely related to thesuppression of HBV cccDNA transcription, and miR-152 may be a factorinvolved in the regulation of the methylation of HBV cccDNA.

Accordingly, miRNAs can directly or indirectly alter HBV replication.The close relationship between miRNAs and HBV-related diseases offers anopportunity to use miRNAs or antagomir in combination therapies totreat, cure, or prevent HBV.

VIII. Pharmaceutical Compositions

Hosts, including but not limited to humans, infected with HBV can betreated by administering to the patient an effective amount of theactive compound or a pharmaceutically acceptable prodrug or salt thereofin the presence of a pharmaceutically acceptable carrier or diluent. Theactive materials can be administered by any appropriate route, forexample, orally, parenterally, intravenously, intradermally,subcutaneously, or topically, in liquid or solid form.

A preferred dose of the compound will be in the range of between about0.01 and about 10 mg/kg, more generally, between about 0.1 and 5 mg/kg,and, preferably, between about 0.5 and about 2 mg/kg, of body weight ofthe recipient per day. The effective dosage range of thepharmaceutically acceptable salts and prodrugs can be calculated basedon the weight of the parent compound to be delivered. If the salt orprodrug exhibits activity in itself, the effective dosage can beestimated as above using the weight of the salt or prodrug, or by othermeans known to those skilled in the art.

The compound is conveniently administered in unit any suitable dosageform, including but not limited to one containing 7 to 600 mg,preferably 70 to 600 mg of active ingredient per unit dosage form. Anoral dosage of 1-400 mg is usually convenient.

The concentration of active compound in the drug composition will dependon absorption, inactivation and excretion rates of the drug as well asother factors known to those of skill in the art. It is to be noted thatdosage values will also vary with the severity of the condition to bealleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat the concentration ranges set forth herein are exemplary only andare not intended to limit the scope or practice of the claimedcomposition. The active ingredient can be administered at once, or canbe divided into a number of smaller doses to be administered at varyingintervals of time.

A preferred mode of administration of the active compound is oral,although for certain patients a sterile injectable form can be given sc,ip or iv. Oral compositions will generally include an inert diluent oran edible carrier. They can be enclosed in gelatin capsules orcompressed into tablets. For the purpose of oral therapeuticadministration, the active compound can be incorporated with excipientsand used in the form of tablets, troches or capsules. Pharmaceuticallycompatible binding agents, and/or adjuvant materials can be included aspart of the composition.

The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a bindersuch as microcrystalline cellulose, gum tragacanth or gelatin; anexcipient such as starch or lactose, a disintegrating agent such asalginic acid, Primogel or corn starch; a lubricant such as magnesiumstearate or Sterotes; a glidant such as colloidal silicon dioxide; asweetening agent such as sucrose or saccharin; or a flavoring agent suchas peppermint, methyl salicylate, or orange flavoring. When the dosageunit form is a capsule, it can contain, in addition to material of theabove type, a liquid carrier such as a fatty oil. In addition, unitdosage forms can contain various other materials that modify thephysical form of the dosage unit, for example, coatings of sugar,shellac, or other enteric agents.

The compound can be administered as a component of an elixir,suspension, syrup, wafer, chewing gum or the like. A syrup can contain,in addition to the active compound(s), sucrose as a sweetening agent andcertain preservatives, dyes and colorings and flavors.

The compound or a pharmaceutically acceptable prodrug or salts thereofcan also be mixed with other active materials that do not impair thedesired action, or with materials that supplement the desired action,such as antibiotics, antifungals, anti-inflammatories or other antiviralcompounds. Solutions or suspensions used for parenteral, intradermal,subcutaneous, or topical application can include the followingcomponents: a sterile diluent such as water for injection, salinesolution, fixed oils, polyethylene glycols, glycerine, propylene glycolor other synthetic solvents; antibacterial agents such as benzyl alcoholor methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents, such as ethylenediaminetetraacetic acid;buffers, such as acetates, citrates or phosphates, and agents for theadjustment of tonicity, such as sodium chloride or dextrose. Theparental preparation can be enclosed in ampoules, disposable syringes ormultiple dose vials made of glass or plastic.

If administered intravenously, preferred carriers are physiologicalsaline or phosphate buffered saline (PBS).

Transdermal Formulations

In some embodiments, the compositions are present in the form oftransdermal formulations, such as that used in the FDA-approved agonistrotigitine transdermal (Neupro patch). Another suitable formulation isdescribed in U.S. Publication No. 20080050424, entitled “TransdermalTherapeutic System for Treating Parkinsonism.” This formulation includesa silicone or acrylate-based adhesive, and can include an additivehaving increased solubility for the active substance, in an amounteffective to increase dissolving capacity of the matrix for the activesubstance.

The transdermal formulations can be single-phase matrices that include abacking layer, an active substance-containing self-adhesive matrix, anda protective film to be removed prior to use. More complicatedembodiments contain multiple-layer matrices that may also containnon-adhesive layers and control membranes. If a polyacrylate adhesive isused, it can be crosslinked with multivalent metal ions such as zinc,calcium, aluminum, or titanium ions, such as aluminum acetylacetonateand titanium acetylacetonate.

When silicone adhesives are used, they are typicallypolydimethylsiloxanes. However, other organic residues such as, forexample, ethyl groups or phenyl groups may in principle be presentinstead of the methyl groups. Because the active compounds are amines,it may be advantageous to use amine-resistant adhesives. Representativeamine-resistant adhesives are described, for example, in EP 0 180 377.

Representative acrylate-based polymer adhesives include acrylic acid,acrylamide, hexyl acrylate, 2-ethylhexylacrylate, hydroxyethylacrylate,octylacrylate, butylacrylate, methylacrylate, glycidylacrylate,methacrylic acid, methacrylamide, hexylmethacrylate,2-ethylhexylmethacrylate, octylmethacrylate, methylmethacrylate,glycidylmethacrylate, vinylacetate, vinylpyrrolidone, and combinationsthereof.

The adhesive must have a suitable dissolving capacity for the activesubstance, and the active substance most be able to move within thematrix, and be able to cross through the contact surface to the skin.Those of skill in the art can readily formulate a transdermalformulation with appropriate transdermal transport of the activesubstance.

Certain pharmaceutically acceptable salts tend to be more preferred foruse in transdermal formulations, because they can help the activesubstance pass the barrier of the stratum corneum. Examples includefatty acid salts, such as stearic acid and oleic acid salts. Oleate andstearate salts are relatively lipophilic, and can even act as apermeation enhancer in the skin.

Permeation enhancers can also be used. Representative permeationenhancers include fatty alcohols, fatty acids, fatty acid esters, fattyacid amides, glycerol or its fatty acid esters, N-methylpyrrolidone,terpenes such as limonene, alpha-pinene, alpha-terpineol, carvone,carveol, limonene oxide, pinene oxide, and 1,8-eucalyptol.

The patches can generally be prepared by dissolving or suspending theactive agent in ethanol or in another suitable organic solvent, thenadding the adhesive solution with stirring. Additional auxiliarysubstances can be added either to the adhesive solution, the activesubstance solution or to the active substance-containing adhesivesolution. The solution can then be coated onto a suitable sheet, thesolvents removed, a backing layer laminated onto the matrix layer, andpatches punched out of the total laminate.

Nanoparticulate Compositions

The compounds described herein can also be administered in the form ofnanoparticulate compositions.

In one embodiment, the controlled release nanoparticulate formulationscomprise a nanoparticulate active agent to be administered and arate-controlling polymer which functions to prolong the release of theagent following administration. In this embodiment, the compositions canrelease the active agent, following administration, for a time periodranging from about 2 to about 24 hours or up to 30 days or longer.Representative controlled release formulations including ananoparticulate form of the active agent are described, for example, inU.S. Pat. No. 8,293,277.

Nanoparticulate compositions comprise particles of the active agentsdescribed herein, having a non-crosslinked surface stabilizer adsorbedonto, or associated with, their surface.

The average particle size of the nanoparticulates is typically less thanabout 800 nm, more typically less than about 600 nm, still moretypically less than about 400 nm, less than about 300 nm, less thanabout 250 nm, less than about 100 nm, or less than about 50 nm. In oneaspect of this embodiment, at least 50% of the particles of active agenthave an average particle size of less than about 800, 600, 400, 300,250, 100, or 50 nm, respectively, when measured by light scatteringtechniques.

A variety of surface stabilizers are typically used with nanoparticulatecompositions to prevent the particles from clumping or aggregating.Representative surface stabilizers include, but are not limited to,gelatin, lecithin, dextran, gum acacia, cholesterol, tragacanth, stearicacid, benzalkonium chloride, calcium stearate, glycerol monostearate,cetostearyl alcohol, cetomacrogol emulsifying wax, sorbitan esters,polyoxyethylene alkyl ethers, polyoxyethylene castor oil derivatives,polyoxyethylene sorbitan fatty acid esters, polyethylene glycols,polyoxyethylene stearates, colloidal silicon dioxide, phosphates, sodiumdodecylsulfate, carboxymethylcellulose calcium, carboxymethylcellulosesodium, methylcellulose, hydroxyethyl cellulose, hydroxypropylcellulose,hydroxypropylmethyl-cellulose phthalate, noncrystalline cellulose,magnesium aluminum silicate, triethanolamine, polyvinyl alcohol,polyvinylpyrrolidone, tyloxapol, poloxamers, poloxamines, poloxamine908, dialkylesters of sodium sulfosuccinic acid, sodium lauryl sulfate,an alkyl aryl polyether sulfonate, a mixture of sucrose stearate andsucrose distearate, p-isononylphenoxypoly-(glycidol), SA9OHCO,decanoyl-N-methylglucamide, n-decyl-D-glucopyranoside,n-decyl-D-maltopyranoside, n-dodecyl-D-glucopyranoside,n-dodecyl-D-maltoside, heptanoyl-N-methylglucamide,n-heptyl-D-glucopyranoside, n-heptyl-D-thioglucoside,n-hexyl-D-glucopyranoside, nonanoyl-N-methylglucamide,n-nonyl-D-glucopyranoside, octanoyl-N-methylglucamide,n-octyl-D-glucopyranoside, and octyl-D-thioglucopyranoside. Lysozymescan also be used as surface stabilizers for nanoparticulatecompositions. Certain nanoparticles such as poly(lactic-co-glycolicacid) (PLGA)-nanoparticles are known to target the liver when given byintravenous (IV) or subcutaneously (SQ).

Because HBV causes damage to, and are present in the liver, in oneembodiment, the nanoparticles or other drug delivery vehicles aretargeted to the liver. One such type of liver-targeted drug deliveryvehicle is described in Park, et al., Mol Imaging. February 2011; 10(1):69-77, and uses Glypican-3 (GPC3) as a molecular target. Park taughtusing this target for hepatocellular carcinoma (HCC), a primary livercancer frequently caused by chronic persistent hepatitis.

In one aspect of this embodiment, this drug delivery vehicle is alsoused to target therapeutics to the liver to treat viral infections.Further, since the compounds described herein have indirect anti-canceruses, this type of system can target the compounds to the liver andtreat liver cancers or reverse the cancer. GPC3 is a heparan sulfateproteoglycan that is not expressed in normal adult tissues, butsignificantly over-expressed in up to 80% of human HCC's. GPC3 can betargeted, for example, using antibody-mediated targeting and binding(See Hsu, et al., Cancer Res. 1997; 57:5179-84).

Another type of drug delivery system for targeting the liver isdescribed in U.S. Pat. No. 7,304,045. The '045 patent discloses adual-particle tumor or cancer targeting system that includes a firstligand-mediated targeting nanoparticle conjugated with galactosamine,with the ligand being on a target cell. The first nanoparticle includespoly(γ-glutamic acid)/poly(lactide) block copolymers and n antiviralcompound, which in this case is a compound described herein, and in the'045 patent, was ganciclovir. A second nanoparticle includespoly(γ-glutamic acid)/poly(lactide) block copolymers, an endothelialcell-specific promoter, and a (herpes-simplex-virus)-(thymidine kinase)gene constructed plasmid, and provides enhanced permeability andretention-mediated targeting. The first and said second nanoparticlesare mixed in a solution configured for delivering to the liver. When thedisorder to be treated is a liver tumor or cancer, the delivery can bedirectly to, or adjacent to, the liver tumor or cancer.

Representative rate controlling polymers into which the nanoparticlescan be formulated include chitosan, polyethylene oxide (PEO), polyvinylacetate phthalate, gum arabic, agar, guar gum, cereal gums, dextran,casein, gelatin, pectin, carrageenan, waxes, shellac, hydrogenatedvegetable oils, polyvinylpyrrolidone, hydroxypropyl cellulose (HPC),hydroxyethyl cellulose (HEC), hydroxypropyl methylcelluose (HPMC),sodium carboxymethylcellulose (CMC), poly(ethylene) oxide, alkylcellulose, ethyl cellulose, methyl cellulose, carboxymethyl cellulose,hydrophilic cellulose derivatives, polyethylene glycol,polyvinylpyrrolidone, cellulose acetate, cellulose acetate butyrate,cellulose acetate phthalate, cellulose acetate trimellitate, polyvinylacetate phthalate, hydroxypropylmethyl cellulose phthalate,hydroxypropylmethyl cellulose acetate succinate, polyvinylacetaldiethylamino acetate, poly(alkylmethacrylate), poly(vinylacetate), polymers derived from acrylic or methacrylic acid and theirrespective esters, and copolymers derived from acrylic or methacrylicacid and their respective esters.

Methods of making nanoparticulate compositions are described, forexample, in U.S. Pat. Nos. 5,518,187 and 5,862,999, both for “Method ofGrinding Pharmaceutical Substances;” U.S. Pat. No. 5,718,388, for“Continuous Method of Grinding Pharmaceutical Substances;” and U.S. Pat.No. 5,510,118 for “Process of Preparing Therapeutic CompositionsContaining Nanoparticles.”

Nanoparticulate compositions are also described, for example, in U.S.Pat. No. 5,298,262 for “Use of Ionic Cloud Point Modifiers to PreventParticle Aggregation During Sterilization;” U.S. Pat. No. 5,302,401 for“Method to Reduce Particle Size Growth During Lyophilization;” U.S. Pat.No. 5,318,767 for “X-Ray Contrast Compositions Useful in MedicalImaging;” U.S. Pat. No. 5,326,552 for “Novel Formulation ForNanoparticulate X-Ray Blood Pool Contrast Agents Using High MolecularWeight Non-ionic Surfactants;” U.S. Pat. No. 5,328,404 for “Method ofX-Ray Imaging Using Iodinated Aromatic Propanedioates;” U.S. Pat. No.5,336,507 for “Use of Charged Phospholipids to Reduce NanoparticleAggregation;” U.S. Pat. No. 5,340,564 for Formulations Comprising Olin10-G to Prevent Particle Aggregation and Increase Stability;” U.S. Pat.No. 5,346,702 for “Use of Non-Ionic Cloud Point Modifiers to MinimizeNanoparticulate Aggregation During Sterilization;” U.S. Pat. No.5,349,957 for “Preparation and Magnetic Properties of Very SmallMagnetic-Dextran Particles;” U.S. Pat. No. 5,352,459 for “Use ofPurified Surface Modifiers to Prevent Particle Aggregation DuringSterilization;” U.S. Pat. Nos. 5,399,363 and 5,494,683, both for“Surface Modified Anticancer Nanoparticles;” U.S. Pat. No. 5,401,492 for“Water Insoluble Non-Magnetic Manganese Particles as Magnetic ResonanceEnhancement Agents;” U.S. Pat. No. 5,429,824 for “Use of Tyloxapol as aNanoparticulate Stabilizer;” U.S. Pat. No. 5,447,710 for “Method forMaking Nanoparticulate X-Ray Blood Pool Contrast Agents Using HighMolecular Weight Non-ionic Surfactants;” U.S. Pat. No. 5,451,393 for“X-Ray Contrast Compositions Useful in Medical Imaging;” U.S. Pat. No.5,466,440 for “Formulations of Oral Gastrointestinal Diagnostic X-RayContrast Agents in Combination with Pharmaceutically Acceptable Clays;”U.S. Pat. No. 5,470,583 for “Method of Preparing NanoparticleCompositions Containing Charged Phospholipids to Reduce Aggregation;”U.S. Pat. No. 5,472,683 for “Nanoparticulate Diagnostic Mixed CarbamicAnhydrides as X-Ray Contrast Agents for Blood Pool and Lymphatic SystemImaging;” U.S. Pat. No. 5,500,204 for “Nanoparticulate Diagnostic Dimersas X-Ray Contrast Agents for Blood Pool and Lymphatic System Imaging;”U.S. Pat. No. 5,518,738 for “Nanoparticulate NSAID Formulations;” U.S.Pat. No. 5,521,218 for “Nanoparticulate Iododipamide Derivatives for Useas X-Ray Contrast Agents;” U.S. Pat. No. 5,525,328 for “NanoparticulateDiagnostic Diatrizoxy Ester X-Ray Contrast Agents for Blood Pool andLymphatic System Imaging;” U.S. Pat. No. 5,543,133 for “Process ofPreparing X-Ray Contrast Compositions Containing Nanoparticles;” U.S.Pat. No. 5,552,160 for “Surface Modified NSAID Nanoparticles;” U.S. Pat.No. 5,560,931 for “Formulations of Compounds as NanoparticulateDispersions in Digestible Oils or Fatty Acids;” U.S. Pat. No. 5,565,188for “Polyalkylene Block Copolymers as Surface Modifiers forNanoparticles;” U.S. Pat. No. 5,569,448 for “Sulfated Non-ionic BlockCopolymer Surfactant as Stabilizer Coatings for NanoparticleCompositions;” U.S. Pat. No. 5,571,536 for “Formulations of Compounds asNanoparticulate Dispersions in Digestible Oils or Fatty Acids;” U.S.Pat. No. 5,573,749 for “Nanoparticulate Diagnostic Mixed CarboxylicAnydrides as X-Ray Contrast Agents for Blood Pool and Lymphatic SystemImaging;” U.S. Pat. No. 5,573,750 for “Diagnostic Imaging X-Ray ContrastAgents;” U.S. Pat. No. 5,573,783 for “Redispersible Nanoparticulate FilmMatrices With Protective Overcoats;” U.S. Pat. No. 5,580,579 for“Site-specific Adhesion Within the GI Tract Using NanoparticlesStabilized by High Molecular Weight, Linear Poly(ethylene Oxide)Polymers;” U.S. Pat. No. 5,585,108 for “Formulations of OralGastrointestinal Therapeutic Agents in Combination with PharmaceuticallyAcceptable Clays;” U.S. Pat. No. 5,587,143 for “Butylene Oxide-EthyleneOxide Block Copolymers Surfactants as Stabilizer Coatings forNanoparticulate Compositions;” U.S. Pat. No. 5,591,456 for “MilledNaproxen with Hydroxypropyl Cellulose as Dispersion Stabilizer;” U.S.Pat. No. 5,593,657 for “Novel Barium Salt Formulations Stabilized byNon-ionic and Anionic Stabilizers;” U.S. Pat. No. 5,622,938 for “SugarBased Surfactant for Nanocrystals;” U.S. Pat. No. 5,628,981 for“Improved Formulations of Oral Gastrointestinal Diagnostic X-RayContrast Agents and Oral Gastrointestinal Therapeutic Agents;” U.S. Pat.No. 5,643,552 for “Nanoparticulate Diagnostic Mixed Carbonic Anhydridesas X-Ray Contrast Agents for Blood Pool and Lymphatic System Imaging;”U.S. Pat. No. 5,718,388 for “Continuous Method of GrindingPharmaceutical Substances;” U.S. Pat. No. 5,718,919 for “NanoparticlesContaining the R(−)Enantiomer of Ibuprofen;” U.S. Pat. No. 5,747,001 for“Aerosols Containing Beclomethasone Nanoparticle Dispersions;” U.S. Pat.No. 5,834,025 for “Reduction of Intravenously AdministeredNanoparticulate Formulation Induced Adverse Physiological Reactions;”U.S. Pat. No. 6,045,829 “Nanocrystalline Formulations of HumanImmunodeficiency Virus (HIV) Protease Inhibitors Using CellulosicSurface Stabilizers;” U.S. Pat. No. 6,068,858 for “Methods of MakingNanocrystalline Formulations of Human Immunodeficiency Virus (HIV)Protease Inhibitors Using Cellulosic Surface Stabilizers;” U.S. Pat. No.6,153,225 for “Injectable Formulations of Nanoparticulate Naproxen;”U.S. Pat. No. 6,165,506 for “New Solid Dose Form of NanoparticulateNaproxen;” U.S. Pat. No. 6,221,400 for “Methods of Treating MammalsUsing Nanocrystalline Formulations of Human Immunodeficiency Virus (HIV)Protease Inhibitors;” U.S. Pat. No. 6,264,922 for “Nebulized AerosolsContaining Nanoparticle Dispersions;” U.S. Pat. No. 6,267,989 for“Methods for Preventing Crystal Growth and Particle Aggregation inNanoparticle Compositions;” U.S. Pat. No. 6,270,806 for “Use ofPEG-Derivatized Lipids as Surface Stabilizers for NanoparticulateCompositions;” U.S. Pat. No. 6,316,029 for “Rapidly Disintegrating SolidOral Dosage Form,” U.S. Pat. No. 6,375,986 for “Solid DoseNanoparticulate Compositions Comprising a Synergistic Combination of aPolymeric Surface Stabilizer and Dioctyl Sodium Sulfosuccinate;” U.S.Pat. No. 6,428,814 for “Bioadhesive nanoparticulate compositions havingcationic surface stabilizers;” U.S. Pat. No. 6,431,478 for “Small ScaleMill;” and U.S. Pat. No. 6,432,381 for “Methods for targeting drugdelivery to the upper and/or lower gastrointestinal tract,” all of whichare specifically incorporated by reference. In addition, U.S. PatentApplication No. 20020012675 A1, published on Jan. 31, 2002, for“Controlled Release Nanoparticulate Compositions,” describesnanoparticulate compositions, and is specifically incorporated byreference.

The nanoparticle formulations including the compounds described herein,and also in the form of a prodrug or a salt, can be used to treat orprevent infections by hepatitis B virus.

Amorphous small particle compositions are described, for example, inU.S. Pat. No. 4,783,484 for “Particulate Composition and Use Thereof asAntimicrobial Agent;” U.S. Pat. No. 4,826,689 for “Method for MakingUniformly Sized Particles from Water-Insoluble Organic Compounds;” U.S.Pat. No. 4,997,454 for “Method for Making Uniformly-Sized Particles FromInsoluble Compounds;” U.S. Pat. No. 5,741,522 for “Ultrasmall,Non-aggregated Porous Particles of Uniform Size for Entrapping GasBubbles Within and Methods;” and U.S. Pat. No. 5,776,496, for“Ultrasmall Porous Particles for Enhancing Ultrasound Back Scatter.”

Controlled Release Formulations

In a preferred embodiment, the active compounds are prepared withcarriers that will protect the compound against rapid elimination fromthe body, such as a controlled release formulation, including but notlimited to implants and microencapsulated delivery systems.Biodegradable, biocompatible polymers can be used, such as ethylenevinyl acetate, polyanhydrides, polyglycolic acid, collagen,polyorthoesters and polylactic acid. For example, enterically coatedcompounds can be used to protect cleavage by stomach acid. Methods forpreparation of such formulations will be apparent to those skilled inthe art. Suitable materials can also be obtained commercially.

Liposomal suspensions (including but not limited to liposomes targetedto infected cells with monoclonal antibodies to viral antigens) are alsopreferred as pharmaceutically acceptable carriers. These can be preparedaccording to methods known to those skilled in the art, for example, asdescribed in U.S. Pat. No. 4,522,811 (incorporated by reference). Forexample, liposome formulations can be prepared by dissolving appropriatelipid(s) (such as stearoyl phosphatidyl ethanolamine, stearoylphosphatidyl choline, arachadoyl phosphatidyl choline, and cholesterol)in an inorganic solvent that is then evaporated, leaving behind a thinfilm of dried lipid on the surface of the container. An aqueous solutionof the active compound is then introduced into the container. Thecontainer is then swirled by hand to free lipid material from the sidesof the container and to disperse lipid aggregates, thereby forming theliposomal suspension.

The terms used in describing the invention are commonly used and knownto those skilled in the art. As used herein, the following abbreviationshave the indicated meanings:

-   ACN Acetonitrile-   Boc₂O Di-tert-butyl dicarbonate-   CDI carbonyldiimidazole-   DCC N,N′-dicyclohexylcarbodiimide-   DCM dichloromethane-   DIPEA diisopropyl ethyl amine (Hünig's base)-   DMAP 4-Dimethylaminopyridine-   DMF N,N-dimethylformamide-   DMSO dimethylsulfoxide-   EDC 1-ethyl-3-(3-dimethyllaminopropyl)carbodiimide hydrochloride-   EtOAc ethyl acetate-   h hour-   HATU    1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide    hexafluorophosphate-   M molar-   min minute-   rt or RT room temperature-   TFA trifluoroacetic acid-   THF tetrahydrofuran-   DMA Dimethylacetamide

IX. General Methods for Preparing Active Compounds

Methods for the facile preparation of active compounds are known in theart and result from the selective combination of known methods. Thecompounds disclosed herein can be prepared as described in detail below,or by other methods known to those skilled in the art. It will beunderstood by one of ordinary skill in the art that variations of detailcan be made without departing from the spirit and in no way limiting thescope of the present invention.

The various reaction schemes are summarized below.

Scheme 1 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound A.Scheme 2 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, an alternate syntheticapproach to compound B.Scheme 3 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound C.Scheme 4 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound D.Scheme 5 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound E.Scheme 6 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound F and G.Scheme 7 is a non-limiting example of the synthesis of active compoundsof the present invention, and in particular, a synthetic approach tocompound H.

Compounds of formula A can be prepared by first selective reaction of ananiline derivative with a carboxylic acid chloride of general formula Iin the presence of an organic base such as Et₃N or DIPEA. IntermediateIII is then reacted with an amine of general formula IV, for example inan organic solvent like CH₂Cl₂, in the presence of an organic base suchas Et₃N.

Compounds of formula B can be prepared by first selective reaction of anamine derivative with a carboxylic acid chloride of general formula V inthe presence of an organic base such as Et₃N or DIPEA. Intermediate VIis then reacted with an amine of general formula IV, for example in anorganic solvent like CH₂Cl₂, in the presence of an organic base such asEt₃N.

The synthesis of compounds of general formula C can be performed asoutlined in Scheme 3. A carboxylic acid of general formula VII can beN-protected, for example, by treatment with Boc₂O in the presence of abase such as NaHCO₃. Intermediate VIII can be coupled with an amine ofgeneral formula II using a peptide coupling reagent like, for example,EDC in the presence of an organic amine base such as DMAP. The resultingcompound of general formula IX can then be deprotected, for example, inthe presence of TFA when Boc was used as a protecting group and thenreacted with a sulfonyl chloride of general formula X in the presence ofan organic amine base such as Et₃N.

The synthesis of compounds of general formula D can be performed asoutlined in Scheme 4. An ester of general formula XI can be reacted withan oxalyl chloride monoalkylester of general formula XII in the presenceof a Lewis acid like, for example, AlCl₃ to give intermediate XIIISelective hydrolysis with an inorganic base like, for example, NaOHfollowed by the coupling of the resulting alpha keto acid XIV with anamine of general formula IV in the presence of a peptide couplingreagent like, for example, CDI provides compounds of general formula XV.Hydrolysis of the ester moiety with an inorganic base like, for example,NaOH followed by the coupling of the resulting carboxyl acid with anamine of general formula II in the presence of a peptide couplingreagent like, for example, HATU in the presence of an organic amine basesuch as DIPEA gives compounds of general formula D.

The synthesis of compounds of general formula E can be performed asoutlined in Scheme 5. A carboxylic acid of general formula XVII can becoupled with an amine of general formula II using a peptide couplingreagent like, for example, HATU in the presence of an organic amine basesuch as DIPEA. Intermediate XVIII can be reacted with an oxalyl chloridemonoalkylester of general formula XII in the presence of a Lewis acidlike, for example, AlCl₃ to give intermediate XIX. Selective hydrolysiswith an inorganic base like, for example, NaOH followed by the couplingof the resulting alpha keto acid XX with an alcohol of general formulaXXI in the presence of a peptide coupling reagent like, for example, DCCin the presence of an organic amine base such as DMAP provides compoundsof general formula E.

The synthesis of compounds of general formula F and G can be performedas outlined in Scheme 6. A carboxylic acid of general formula XXII canbe coupled with an amine of general formula II using a peptide couplingreagent like, for example, HATU in the presence of an organic amine basesuch as DIPEA. Reduction of compounds XXIII using, for example, Zn inthe presence of formic acid leads to amino derivatives of generalformula XXIV which can be either reacted with an oxoacetic acidderivative of general formula XXV in the presence of a peptide couplingreagent like, for example, DCC or with a sulfonyl chloride of generalformula X in the presence of an organic amine base such as Et₃N toafford respectively compounds of general Formulas F and G.

The synthesis of compounds of general formula H can be performed asoutlined in Scheme 7. A bromo derivative of general formula XXVI canundergo a lithium-halogen exchange using an organolithium reagent suchas, for example, n-BuLi and react with a dialkyloxalate like, forexample, diethyl oxalate. The resulting compound can then be hydrolyzedto form the carboxylic acid of general formula XXVIII which can becoupled with an amine of general formula II using a peptide couplingreagent like, for example, HATU in the presence of an organic amine basesuch as DIPEA. Hydrolysis of compound XXIX with an inorganic base like,for example, NaOH followed by the coupling of the resulting alpha ketoacid XXX with an amine of general formula IV in the presence of apeptide coupling reagent like, for example, CDI in the presence of anorganic amine base such as DIPEA provides compounds of general formulaH.

SPECIFIC EXAMPLES

Specific compounds which are representative of this invention wereprepared as per the following examples and reaction sequences; theexamples and the diagrams depicting the reaction sequences are offeredby way of illustration, to aid in the understanding of the invention andshould not be construed to limit in any way the invention set forth inthe claims which follow thereafter. The present compounds can also beused as intermediates in subsequent examples to produce additionalcompounds of the present invention. No attempt has necessarily been madeto optimize the yields obtained in any of the reactions. One skilled inthe art would know how to increase such yields through routinevariations in reaction times, temperatures, solvents and/or reagents.

Anhydrous solvents were purchased from Aldrich Chemical Company, Inc.(Milwaukee, Wis.) and EMD Chemicals Inc. (Gibbstown, N.J.). Reagentswere purchased from commercial sources. Unless noted otherwise, thematerials used in the examples were obtained from readily availablecommercial suppliers or synthesized by standard methods known to oneskilled in the art of chemical synthesis. ¹H and ¹³C NMR spectra weretaken on a Bruker Ascend™ 400 MHz Fourier transform spectrometer at roomtemperature and reported in ppm downfield from internaltetramethylsilane. Deuterium exchange, decoupling experiments or 2D-COSYwere performed to confirm proton assignments. Signal multiplicities arerepresented by s (singlet), d (doublet), dd (doublet of doublets), t(triplet), q (quadruplet), br (broad), bs (broad singlet), m(multiplet). All J-values are in Hz. Mass spectra were determined on aMicromass Platform LC spectrometer using electrospray techniques.Analytic TLC were performed on Sigma-Aldrich® aluminum supported silicagel (25 μm) plates. Column chromatography was carried out on Silica Gelor via reverse-phase high performance liquid chromatography.

Example 1

Ethyl 1,3,5-Trimethylpyrrole-2-carboxylate (2)

Ethyl 3,5-dimethylpyrrole-2-carboxylate (100.0 g, 0.59 mol) was added toa solution of potassium hydroxide (100.6 g, 1.79 mol) in dimethylsulfoxide (1 L) and stirred for 30 min under nitrogen at 0° C. Methyliodide (55.9 mL, 0.89 mol) was then added and the reaction mixture wasallowed to warm up to room temperature and stirred for 4 hours. Thereaction was then extracted with diethyl ether (3×1 L) and the combinedorganic layer were finally washed with water (2×150 mL), dried overNa₂SO₄ and concentrated in vacuo. The residue slowly crystallized toyield ethyl 1,3,5-trimethylpyrrole-2-carboxylate 2 (102.4 g, 0.56 mol,94%) as a yellowish solid. ¹H NMR (400 MHz, CDCl₃) δ: 1.32 (t, 3H), 2.20(s, 3H), 2.30 (s, 3H), 3.75 (s, 3H), 4.22 (q, 2H), 5.75 (s, 1H). MS(ESI): m/z [M+H]⁺ calcd for C₉H₁₄NO₂: 182.2, found: 182.3.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid (6)

To a solution of ethyl 1,3,5-trimethylpyrrole-2-carboxylate (10.0 g,55.2 mmol) in CH₂Cl₂ (250 mL) at 0° C. was added dropwise a solution ofethyl 2-chloro-2-oxo-acetate (9.3 mL, 82.8 mmol) in CH₂Cl₂ (100 mL)followed by AlCl₃ (22.1 g, 165.7 mmol) portion wise. The reactionmixture was then stirred overnight at room temperature and then quenchedwith ice. After addition of water (300 mL), the mixture was filtered oncelite and extracted with CH₂Cl₂ (3×100 mL). The combined organic layerswere washed with a saturated solution of sodium carbonate (250 mL) and asaturated solution of ammonium chloride (250 mL), dried over Na₂SO₄ andconcentrated in vacuo. To the resulting oil were added methanol (100 mL)and a 5% solution of sodium hydroxide (100 mL) and the mixture wasstirred for 15 min at room temperature. After removal of the methanolunder vacuum, the mixture was washed with ethyl acetate (2×100 mL),acidified with a 1N HCl solution (pH=1) and extracted with ethyl acetate(3×100 mL). The combined organic layers were dried over sodium sulfateand concentrated under vacuum. The resulting solid was washed withdiethyl ether (100 mL) and hexanes (100 mL) to yield2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(8.1 g, 32.0 mmol, 58%) as an off-white powder. To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(2.0 g, 7.9 mmol) in DMF (15 mL) and CH₂Cl₂ (10 mL) was added1,1′-carbonyldiimidazole (1.92 g, 11.8 mmol) and propargylamine (0.607mL, 9.5 mmol). After stirring for 2 h at room temperature, the reactionmixture was poured into a saturated solution of ammonium chloride andextracted with CH₂Cl₂ (3×100 mL).

The combined organic layers were dried over sodium sulfate andconcentrated in vacuo to give 5 as a yellowish oil. To the crude ethyl4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrole-2-carboxylate5 dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solutionof sodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted with ethyl acetate(3×50 mL). The combined organic layers were dried over sodium sulfateand concentrated in vacuo. The resulting solid was washed with diethylether (50 mL) and hexanes (50 mL) to yield4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (1.8 g, 6.9 mmol, 87%) as a white powder. ¹H NMR (400 MHz,DMSO-d6) δ 12.77 (s, 1H), 9.14 (t, J=5.7 Hz, 1H), 3.99 (dd, J=5.7, 2.6Hz, 2H), 3.75 (s, 3H), 3.18 (t, J=2.4 Hz, 1H), 2.37 (s, 6H). ¹³C NMR(101 MHz, DMSO) δ 188.0, 167.2, 163.0, 142.8, 129.7, 121.8, 117.5, 80.5,73.8, 33.3, 28.1, 12.3, 12.0. HRMS (ESI): m/z [M+H]⁺ calcd forC₁₃H₁₅N₂O₄: 263.1032, found: 263.1025.

4-[(Propargylamino)(oxo)acetyl]-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(7a)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (750 mg, 2.7 mmol), 3,4-difluoroaniline (410 mg, 3.2 mmol) andDIPEA (746 μL, 4.3 mmol) in DMF (15 mL) was added HATU (1.63 g, 4.3mmol) at room temperature. The mixture was stirred at 50° C. for 3 h. Inorder to reach completion, more 3,4-difluoroaniline (410 mg, 3.2 mmol)was added and the mixture was further stirred overnight at 65° C. Thereaction mixture was then poured into a saturated solution of ammoniumchloride and extracted with ethyl acetate (3×50 mL).

The combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 7a as a white powder (47%, 503mg, 1.4 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.49 (s, 1H), 8.22-8.07(m, 1H), 8.07-7.95 (m, 1H), 7.59-7.43 (m, 1H), 7.33 (q, J=9.4 Hz, 1H),4.21-4.07 (m, 2H), 3.69 (s, 3H), 2.73 (s, 1H), 2.43 (s, 3H), 2.29 (s,3H). ¹³C NMR (101 MHz, Acetone) δ 188.9, 167.9, 162.1, 152.7, 152.6,150.3, 150.2, 149.3, 149.2, 146.9, 146.8, 142.8, 137.9, 137.9, 128.5,124.6, 119.1, 118.9, 118.8, 117.5, 117.5, 117.4, 117.4, 110.7, 110.5,81.4, 73.2, 33.2, 29.7, 12.8, 12.7. ¹⁹F NMR (377 MHz, Acetone-d6) δ−139.8-−140.0 (m), −147.1-−147.2 (m). FIRMS (ESI): m/z [M+H]⁺ calcd forC₁₉H₁₈F₂N₃O₃: 374.1316, found: 374.1309.

4-(2-(((1-cyclopropyl-1H-1,2,3-triazol-4-yl)methyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(8)

A solution of bromocyclopropane (0.4 mL, 3.3 mmol) and sodium azide (430mg, 6.6 mmol) in water (1 mL) was heated under microwave irradiation for30 minutes at 120° C. A solution of compound 7 (0.05 g, 0.1 mmol) inacetonitrile (1 mL) was then added, followed by sodium ascorbate (10 mg,0.05 mmol) and copper sulfate (20 mg, 0.12 mmol). The mixture was heatedfor 30 minutes at 80° C. under microwave irradiation and then pouredinto a saturated solution of ammonium chloride (50 mL). After extractionwith ethyl acetate (3×50 mL), the combined organic layers were driedover sodium sulfate and concentrated in vacuo. The resulting mixture waspurified by flash chromatography (Hexanes/Ethyl acetate=6:4 v/v), togive compound 8 as a white powder (61%, 31 mg, 0.06 mmol). ¹H NMR (400MHz, Acetone-d₆) δ 9.50 (s, 1H), 8.29-8.18 (m, 1H), 8.06-7.94 (m, 1H),7.87 (s, 1H), 7.57-7.48 (m, 1H), 7.39-7.28 (m, 1H), 6.16-5.98 (m, 1H),5.31-5.20 (m, 1H), 5.09-5.00 (m, 2H), 4.58 (d, J=5.9 Hz, 2H), 3.67 (d,J=1.4 Hz, 3H), 2.38 (d, J=1.3 Hz, 3H), 2.24 (d, J=1.3 Hz, 3H). ¹³C NMR(101 MHz, Acetone) δ 187.4, 166.3, 160.3, 150.9, 150.8, 148.5, 148.4,147.5, 147.4, 145.1, 144.9, 144.4, 144.4, 140.9, 136.2, 136.1, 136.1,136.0, 132.7, 126.6, 122.9, 122.5, 118.3, 117.3, 117.1, 117.1, 115.7,115.7, 115.7, 115.7, 108.9, 108.7, 51.9, 34.4, 31.4, 11.1, 11.0. ¹⁹F NMR(377 MHz, Acetone-d₆) δ −139.8-−140.1 (m), −147.0-−147.2 (m). HRMS(ESI): m/z [M+H]+ calcd for C₂₂H₂₃F₂N₆O₃: 457.1800, found: 457.1790.

N-(3-cyano-4-fluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(7b)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (0.1 g, 0.38 mmol) in toluene (5 mL) was added 0.1 mL of SOCl₂ atroom temperature and the mixture was refluxed for 1.5 h. After removalof SOCl₂ in vacuo, the residual oil was solubilized in DMA (5 mL) and3-cyano-4-fluoroaniline (0.1 g, 0.7 mmol) was added. The mixture wasstirred at 100° C. for 3 h and then cooled down to room temperature. Thesolution was then poured into a saturated solution of ammonium chloride(50 mL) and extracted with EtOAc (3×25 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo.

The resulting residue was purified by flash chromatography(Hexanes/EtOAc=6:4 v/v) to yieldN-(3-cyano-4-fluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide7b (48%, 0.7 g, 0.2 mmol). ¹H NMR (400 MHz Acetone-d₆) δ 9.61 (s, 1H),8.37-8.27 (m, 1H), 8.15 (s, 1H), 8.13-8.04 (m, 1H), 7.45 (t, =9.1 Hz,1H), 4.21-4.06 (m, 2H), 3.70 (s, 3H), 2.77-2.70 (m, 1H), 2.43 (s, 3H),2.30 (s, 3H). ¹³C NMR (101 MHz, Acetone-d₆) δ 188.9, 167.8, 162.3, 160.7(d, J=252.7 Hz) 143.0, 138.0 (d, J=3.1 Hz), 128.4 (d, J=8.1 Hz), 128.2,125.5, 125.0, 118.9, 118.6 (d, J=20.9 Hz), 115.3, 102.6 (d, J=16.5 Hz),81.4, 73.3, 33.3, 29.7, 12.9, 12.8. ¹⁹F NMR (377 MHz, Acetone-d₆) δ−115.9 (dd, J=9.6, 4.7 Hz). LCMS (ESI): m/z [M+H]⁺ calcd forC₂₀H₁₉FN₄O₃: 381.1, found: 381.3.

5-(N-((1-Cyclopropyl-1H-1,2,3-triazol-4-yl)methyl)sulfamoyl)-N-(3,4-difluorophenyl)-2-fluorobenzamide(13)

2-fluorobenzoic acid 9 (10.0 g, 71.4 mmol) was added to chlorosulfonicacid (50 mL) at 0° C. and the mixture was stirred at 0° C. for 1 h. Thereaction mixture was then slowly poured onto ice. The precipitate formedwas filtered, rinsed off with water (3×100 mL) and dried under vacuumovernight to yield 5-(chlorosulfonyl)-2-fluorobenzoic acid 10 as abrownish solid (10.5 g, 44.0 mmol). 5-(chlorosulfonyl)-2-fluorobenzoicacid 10 (10.0 g, 41.9 mmol) was added to 60 mL of SOCl₂ at roomtemperature and the mixture was refluxed for 1.5 h. After removal ofSOCl₂ in vacuo, the residual oil was solubilized in toluene (150 mL) and3,4-difluoroaniline (6.5 g, 50.3 mmol) was added. The mixture wasstirred at 100° C. for 4 h and then cooled down to room temperature. Thesolution was then poured into a saturated solution of ammonium chloride(200 mL) and extracted with EtOAc (3×200 mL). The combined organiclayers were dried over sodium sulfate and concentrated in vacuo. Theresulting residue was purified by flash chromatography(Hexanes/EtOAc=6:4 v/v) to yield3-((3,4-difluorophenyl)carbamoyl)-4-fluorobenzene-1-sulfonyl chloride 11(92%, 13.51 g, 38.6 mmol). To a solution of3-((3,4-difluorophenyl)carbamoyl)-4-fluorobenzene-1-sulfonyl chloride 11(10.0 g, 28.6 mmol) in CH₂Cl₂ (200 mL) at 0° C. were addedpropargylamine hydrochloride (3.1 g, 34.3 mmol) and Et₃N (7.8 mL, 57.2mmol). The reaction mixture was stirred overnight at room temperatureand then poured into a saturated solution of ammonium chloride (250 mL).After extraction with CH₂Cl₂ (3×100 mL), the combined organic layerswere dried over sodium sulfate and finally concentrated in vacuo. Theresulting solid was washed with diethyl ether (50 mL) and hexanes (50mL) to yieldN-(3,4-difluorophenyl)-2-fluoro-5-(N-(prop-2-yn-1-yl)sulfamoyl)benzamide12 (74%, 7.8 g, 21.1 mmol) as a white powder. A solution ofbromocyclopropane (0.6 mL, 4.9 mmol) and sodium azide (483 mg, 7.4 mmol)in water (1 mL) was heated under microwave irradiation for 30 minutes at120° C. To this solution were added a solution of compound 12 (0.1 g,0.3 mmol) in acetonitrile (1 mL), sodium ascorbate (10 mg, 0.05 mmol)and copper sulfate (20 mg, 0.12 mmol).

The reaction mixture was then heated for 30 minutes at 80° C. undermicrowave irradiation before being poured into a saturated solution ofammonium chloride (50 mL). After extraction with EtOAc (3×50 mL), thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The resulting residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) giving compound 13 as a white powder (19%, 23mg, 0.05 mmol). ¹H NMR (400 MHz, Acetone-d6) δ 9.92 (s, 1H), 8.25 (dd,J=6.6, 2.5 Hz, 1H), 8.06-7.95 (m, 2H), 7.77 (s, 1H), 7.58-7.52 (m, 1H),7.48 (dd, J=10.1, 8.7 Hz, 1H), 7.37 (dt, J=10.6, 9.0 Hz, 1H), 7.21 (brs,1H), 6.11-5.94 (m, 1H), 5.32-5.16 (m, 1H), 4.99 (dt, =6.0, 1.5 Hz, 2H),4.32 (s, 2H). ¹³C NMR (101 MHz, Acetone-d₆) δ 188.9, 167.9, 162.1,150.2, 149.2 (d, J=12.9 Hz), 146.8 (d, J=13.0 Hz), 142.8, 137.9 (d,J=5.9 Hz), 128.5, 124.6, 120.4-118.2 (m), 117.5 (dd, J=6.0, 3.6 Hz),110.6 (d, J=22.1 Hz), 81.5, 73.3, 33.2, 29.7, 12.8 (d, J=9.6 Hz). ¹⁹FNMR (377 MHz, Acetone-d6) δ −110.6 (s), −139.6-−139.7 (m), −146.1-−146.3(m). HRMS (ESI): m/z [M+H]⁺ calcd for C₁₉H₁₇F₃N₅O₃S: 452.1004, found:452.0999.

N-(3,4-Difluorophenyl)-1-methyl-4-(N-(prop-2-yn-1-yl)sulfamoyl)-1H-pyrrole-2-carboxamide(17)

1-methyl-1H-pyrrole-2-carboxylic acid (10.0 g, 80 mmol) was added tochlorosulfonic acid (50 mL) at 0° C. and the resulting solution wasstirred at 0° C. for 1 h. The reaction mixture was then slowly pouredonto ice. The precipitate formed was then filtered, rinsed with water(3×100 mL) and dried under vacuum overnight to yield4-(chlorosulfonyl)-1-methyl-1H-pyrrole-2-carboxylic acid 15 as abrownish solid (10.7 g, 47.8 mmol).4-(chlorosulfonyl)-1-methyl-1H-pyrrole-2-carboxylic acid 15 (1.0 g, 4.4mmol) was added to SOCl₂ at room temperature and the mixture was heatedat 100° C. for 1.5 h. After removal of thionyl chloride in vacuo, theresidual oil was solubilized in toluene (50 mL) and 3,4-difluoroanilinewas added (650 mg, 5.0 mmol).

The solution was stirred for 48 h at room temperature before beingpoured into a saturated solution of ammonium chloride (50 mL). Thissolution was extracted with ethyl acetate (3×50 mL) and the combinedorganic layers were dried over sodium sulfate. After concentration invacuo, the resulting residue was purified by flash chromatography(Hexanes:EtOAc=7:3 v/v) to yield5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrole-3-sulfonylchloride 16 (775 mg, 2.3 mmol). To a solution of5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrole-3-sulfonylchloride 16 (100 mg, 0.3 mmol) in DMF (5 mL) were added propargylaminehydrochloride (33 mg, 0.4 mmol) and DIPEA (78 μL, 0.5 mmol). Thereaction mixture was stirred overnight at room temperature, poured intoa saturated solution of ammonium chloride (50 mL) and extracted withethyl acetate (3×50 mL). The combined organic layers were dried oversodium sulfate and concentrated in vacuo.

The resulting residue was purified by flash chromatography(Hexanes:EtOAc=7:3 v/v) to yieldN-(3,4-difluorophenyl)-1-methyl-4-(N-(prop-2-yn-1-yl)sulfamoyl)-1H-pyrrole-2-carboxamide17 (77%, 81 mg, 0.2 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.56 (s, 1H),8.05-7.87 (m, 1H), 7.62-7.48 (m, 2H), 7.38-7.25 (m, 1H), 7.22 (d, J=1.9Hz, 1H), 6.57 (s, 1H), 4.03 (s, 3H), 3.79 (s, 2H), 2.70 (t, J=2.5 Hz,1H). ¹³C NMR (101 MHz, Acetone-d₆) δ 160.9, 152.6 (d, J=13.0 Hz), 150.1(d, J=13.2 Hz), 149.1 (d, J=12.7 Hz), 146.7 (d, J=12.8 Hz), 137.9 (dd,J=9.1, 3.0 Hz), 132.2, 128.4, 123.7, 119.7-118.4 (m), 117.8 (dd, J=5.9,3.5 Hz), 114.0, 110.8 (d, J=22.1 Hz), 80.8, 74.5, 38.5, 34.2. ¹⁹F NMR(377 MHz, Acetone-d₆) δ −138.4-−138.7 (m), −145.4-−146.1 (m). HRMS(ESI): m/z [M+H]⁺ calcd for C₁₅H₁₄F₂N₃O₃S: 354.0724, found: 354.0717.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid (20)

Ethyl 1,3,5-trimethylpyrrole-2-carboxylate 2 (2.0 g, 11.0 mmol) wasadded to chlorosulfonic acid (10 mL) at 0° C. and the mixture wasstirred at 0° C. for 1 h then 3 h at room temperature. The reactionmixture was then slowly poured onto ice. The mixture was basified with a5% solution of sodium hydroxide (pH>7) and extracted with ethyl acetate(3×100 mL). The organic layers were combined, dried over Na₂SO₄ andconcentrated in vacuo to yield 18 a crude dark brown solid (69%, 2.1 g,7.5 mmol). To the crude ethyl4-(chlorosulfonyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylate intermediate18 solubilized in DMF (10 mL) were added propargylamine (0.72 mL, 11.3mmol) and triethylamine (3.1 mL, 22.5 mmol). The mixture was stirred atroom temperature for 2 h and poured into a saturated solution ofammonium chloride (50 mL). After extraction with EtOAc (3×50 mL), thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The resulting oil was diluted in methanol (5 mL) and a 5%solution of sodium hydroxide (15 mL) was added. The mixture was stirredovernight at 60° C. After removal of the methanol in vacuo, the mixturewas washed with ethyl acetate (2×50 mL), acidified with a 1N HClsolution (pH=1) and extracted with ethyl acetate (3×50 mL). The combinedorganic layers were dried over sodium sulfate and concentrated in vacuo.The resulting solid was washed with diethyl ether (20 mL) and hexanes(20 mL) to yield1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid 20 (28%, 0.8 g, 0.3 mmol) as an off-white powder. ¹H NMR (400 MHz,DMSO-d₆) δ 12.70 (s, 1H), 7.64 (t, J=5.9 Hz, 1H), 3.73 (s, 3H), 3.58(dd, J=6.0, 2.6 Hz, 2H), 3.04 (t, J=2.5 Hz, 1H), 2.43 (s, 3H), 2.41 (s,3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 163.0, 138.9, 127.6, 120.8, 118.1,80.1, 74.3, 33.4, 31.8, 12.0, 11.5.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(21)

To a solution of1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid 20 (0.1 g, 0.3 mmol), 3,4-difluoroaniline (96 mg, 7.4 mmol) andDIPEA (746 μL, 1.1 mmol) in DMF (15 mL) was added HATU (0.211 g, 0.5mmol) at room temperature. The mixture was stirred at 50° C. overnight.The reaction mixture was then poured into a saturated solution ofammonium chloride (50 mL) and extracted with ethyl acetate (3×50 mL).The combined organic layers were dried over sodium sulfate, andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 21 as a white powder (36%, 51mg, 0.1 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.44 (s, 1H), 8.24-7.80(m, 1H), 7.74-7.44 (m, 1H), 7.40-7.22 (m, 1H), 6.51 (d, J=6.3 Hz, 1H),3.77-3.70 (m, 2H), 3.68 (s, 3H), 2.68-2.60 (m, 1H), 2.51 (s, 3H), 2.37(s, 3H). ¹³C NMR (101 MHz, Acetone-d₆) δ 160.2, 150.9 (d, J=13.1 Hz),148.4 (d, J=13.0 Hz), 147.5 (d, J=12.9 Hz), 145.1 (d, J=12.6 Hz), 137.1,126.1, 120.7, 117.2 (d, J=18.1 Hz), 117.0, 115.8 (d, J=3.9 Hz), 108.9(d, J=22.2 Hz), 79.1, 72.3, 31.7, 10.4 (d, J=32.3 Hz). ¹⁹F NMR (377 MHz,Acetone-d₆) δ −139.8-−139.9 (m), −147.0-−147.1 (m). HRMS (ESI): m/z[M+H]⁺ calcd for C₁₇H₁₈F₂N₃O₃S: 382.1037, found: 382.1027.

2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid (24)

To a solution of 1-methyl-1H-pyrrole-2-carboxylic acid (4 g, 32 mmol),3,4-difluoroaniline (6.2 g, 48 mmol) and DIPEA (13 mL, 96 mmol) in DMF(150 mL) was added HATU (18.2 g, 48 mmol) at room temperature. Themixture was heated at 65° C. overnight and poured into a saturatedsolution of ammonium chloride. After extraction with ethyl acetate (3×50mL), the combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The resulting mixture was purified by flashchromatography (hexanes:EtOAc=6:4 v/v), to give compound 22 as a whitepowder (82%, 6.2 g, 26.2 mmol).

To a solution ofN-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide 22 (3 g, 12.7mmol) in CH₂Cl₂ (150 mL) was added dropwise, at 0° C., a solution ofethyl 2-chloro-2-oxo-acetate (2.12 mL, 19.1 mmol) in CH₂Cl₂ (20 mL),followed by AlCl₃ (5.1 g, 38.1 mmol) portion wise. The resulting mixturewas stirred overnight at room temperature and then poured onto crushedice. After addition of water (300 mL), the mixture was filtered onCelite and the aqueous layer was extracted with CH₂Cl₂ (3×100 mL). Thecombined organic layers were washed with a saturated solution of sodiumcarbonate (100 mL), a saturated solution of ammonium chloride (100 mL),dried over Na₂SO₄ and concentrated in vacuo. The resulting oil wasdiluted in methanol/THF (v/v=1/2, 30 mL) and a 5% solution of sodiumhydroxide (30 mL) was added. After being stirred for 15 min at roomtemperature, the mixture was concentrated in vacuo. The mixture waswashed with ethyl acetate (2×50 mL), acidified with a 1N HCl solution(pH=1) and extracted with ethyl acetate (3×50 mL). The combined organiclayers were dried over sodium sulfate, and concentrated in vacuo. Theresulting solid was washed with diethyl ether (20 mL) and hexanes (20mL) to yield2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (3.48 g, 11.3 mmol) as an off-white powder. ¹H NMR (400 MHz,DMSO-d₆) δ 10.28 (s, 1H), 8.02 (d, J=1.9 Hz, 1H), 7.94-7.81 (m, 1H),7.60 (d, J=1.9 Hz, 1H), 7.56-7.47 (m, 1H), 7.44-7.24 (m, 1H), 3.96 (s,3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 181.1, 165.3, 159.5, 150.5 (d, J=13.1Hz), 148.0 (d, J=13.1 Hz), 147.0 (d, 0.1=12.5 Hz), 144.6 (d, =12.6 Hz),136.6, 136.4 (dd, 0.1=9.2, 2.8 Hz), 127.9, 119.0, 117.7 (d, J=17.7 Hz),116.8 (dd, J=5.8, 3.2 Hz), 114.9, 109.4 (d, J=21.7 Hz), 37.7. ¹⁹F NMR(377 MHz, DMSO-d₆) δ −138.0-−138.2 (m), −145.5-−145.6 (m).

4-(2-(allylamino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide(25)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol) in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and allylamine (29 μL, 0.4mmol) at room temperature. The mixture was stirred for 2 h at roomtemperature and poured into a saturated solution of ammonium chloride.After extraction with CH₂Cl₂ (3×20 mL), the combined organic layers weredried over sodium sulfate and concentrated in vacuo. The resultingresidue was purified by flash chromatography (Hexanes:EtOAc=7:3 v/v) toafford4-(2-(allylamino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide25 as an off-white powder (81%, 91 mg, 0.2 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.60 (s, 1H), 8.23 (s, 1H), 8.14 (s, 1H), 8.04-7.90 (m,1H), 7.58 (s, 1H), 7.57-7.49 (m, 1H), 7.30 (q, J=9.6 Hz, 1H), 6.25-5.68(m, 1H), 5.22 (d, J=16.8 Hz, 1H), 5.11 (dt, J=10.3, 1.4 Hz, 1H), 4.06(s, 3H), 3.99-3.92 (m, 2H). ¹³C NMR (101 MHz, Acetone-d₆) δ 182.7,163.7, 161.1, 152.6 (d, J=13.1 Hz), 150.2 (d, J=12.9 Hz), 149.1 (d,J=12.8 Hz), 146.7 (d, 0.1=12.9 Hz), 138.9, 137.9, 136.2, 129.0, 120.9,118.8 (d, 0.1=18.0 Hz), 117.9-117.4 (m), 117.0, 116.6, 110.7 (d, J=22.1Hz), 42.9, 38.7. ¹⁹F NMR (377 MHz, Acetone-d₆) δ −138.5-−138.8 (m),−145.9-−146.2 (m). HRMS (ESI): m/z [M+H]⁺ calcd for C₁₇H₁₆F₂N₃O₃:348.1260, found: 348.1158.

N-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxamide(26)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol) in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and thiazole-2-amine (33 mg,0.4 mmol) at room temperature. The mixture was stirred for 2 h at roomtemperature and poured into a saturated solution of ammonium chloride.After extraction with CH₂Cl₂ (3×20 mL), the combined organic layers weredried over sodium sulfate and concentrated in vacuo. The resultingresidue was purified by flash chromatography (Hexanes:EtOAc=7:3 v/v) toaffordN-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxamide26 as an brownish powder (79%, 101 mg, 0.2 mmol). ¹H NMR (400 MHz,DMSO-d₆) δ 12.74 (s, 1H), 10.33 (s, 1H), 8.20 (s, 1H), 7.94-7.83 (m,1H), 7.69 (s, 1H), 7.60 (d, J=3.6 Hz, 1H), 7.56-7.49 (m, 1H), 7.48-7.38(m, 1H), 3.98 (s, 3H). ¹³C NMR (101 MHz, DMSO-d₆) δ 180.6, 162.5, 159.5,150.5 (d, J=12.9 Hz), 148.1 (d, J=12.5 Hz), 138.6, 137.2, 136.4 (d,J=12.0 Hz), 127.9, 118.7, 117.8 (d, J=17.6 Hz), 116.8, 109.5 (d, J=21.7Hz), 60.2, 37.8, 17.9 (d, J=672.1 Hz). ¹⁹F NMR (377 MHz, DMSO-d₆) δ−137.3-−137.4 (m), −144.5-−144.7 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₁₇H₁₃F₂N₄O₃S: 391.0676, found: 391.0669.

4-(2-((cyclopentyloxy)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide(27)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol) in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) andO-cyclopentylhydroxylamine hydrochloride (44 mg, 0.4 mmol) at roomtemperature. The mixture was stirred for 2 h at room temperature andpoured into a saturated solution of ammonium chloride. After extractionwith CH₂Cl₂ (3×20 mL), the combined organic layers were dried oversodium sulfate and concentrated in vacuo. The resulting residue waspurified by flash chromatography (Hexane:EtOAc=7:3 v/v) to give4-(2-((cyclopentyloxy)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide27 as a brownish powder (26%, 33 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 10.83 (s, 1H), 9.63 (s, 1H), 8.14 (d, J=1.7 Hz, 1H),8.03-7.90 (m, 1H), 7.65-7.46 (m, 2H), 7.40-7.20 (m, 1H), 4.69-4.57 (m,1H), 4.06 (s, 3H), 1.95-1.85 (m, 2H), 1.79-1.65 (m, 4H), 1.57 (s, 2H).¹³C NMR (101 MHz, Acetone-d₆) δ 181.0, 161.2 (d, J=178.9 Hz), 159.7,159.3, 150.8 (d, J=13.2 Hz), 148.3 (d, J=13.2 Hz), 147.3 (d, J=12.8 Hz),144.9 (d, J=12.7 Hz), 136.8, 136.2 (dd, J=9.1, 3.0 Hz), 127.4, 119.2,117.0 (d, J=18.0 Hz), 116.5-115.7 (m), 114.5, 109.1 (d, J=22.2 Hz),87.4, 87.0, 37.0, 30.9, 23.4. ¹⁹F NMR (377 MHz, Acetone-d₆) δ−138.6-−138.7 (m), −146.0-−146.1 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₁₉H₂₀F₂N₃O₄: 392.1422, found: 392.1415.

N-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxamide(28)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol) in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and aniline (35 μL, 0.4mmol) at room temperature. The mixture was stirred for 2 h at roomtemperature and poured into a saturated solution of ammonium chloride.After extraction with CH₂Cl₂ (3×20 mL), the combined organic layers weredried over sodium sulfate and concentrated in vacuo. The resultingresidue was purified by flash chromatography (Hexanes:EtOAc=7:3 v/v) toaffordN-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxamide28 as a white powder (32%, 40 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.79 (s, 1H), 9.64 (s, 1H), 8.30 (s, 1H), 8.04-7.94 (m,1H), 7.93 (s, 1H), 7.91 (s, 1H), 7.63 (d, J=1.8 Hz, 1H), 7.60-7.52 (m,1H), 7.44-7.37 (m, 2H), 7.36-7.27 (m, 1H), 7.22-7.14 (m, 1H), 4.10 (s,3H). ¹³C NMR (101 MHz, Acetone-d₆) δ 182.4, 162.0, 161.1, 152.6 (d,J=13.0 Hz), 150.2 (d, J=13.2 Hz), 149.1 (d, J=12.5 Hz), 146.7 (d, J=12.8Hz), 139.7, 139.1, 138.0 (dd, J=9.2, 2.9 Hz), 130.6, 129.2, 126.3,121.8, 120.5, 118.9 (d, J=17.9 Hz), 117.8 (dd, J=5.9, 3.5 Hz), 116.7,110.9 (d, J=22.3 Hz), 38.8. ¹⁹F NMR (377 MHz, Acetone-d₆) δ−138.6-−138.9 (m), −146.1-−146.2 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₂₀H₁₆F₂N₃O₃: 384.116, found: 384.1153.

N-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxamide(29)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol), in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and benzylamine (42 μL, 0.4mmol) at room temperature. The mixture was stirred for 2 h at roomtemperature and poured into a saturated solution of ammonium chloride.After extraction with CH₂Cl₂ (3×20 mL), the combined organic layers weredried over sodium sulfate and concentrated in vacuo. The resultingresidue was purified by flash chromatography (Hexane:EtOAc=7:3 v/v) toaffordN-(3,4-difluorophenyl)-1-methyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxamide29 as a white powder (99%, 126 mg, 0.2 mmol). ¹³C NMR (101 MHz,Acetone-d₆) δ 182.7, 163.9, 161.1, 159.9, 152.6 (d, J=13.0 Hz), 150.1(d, J=13.3 Hz), 149.1 (d, J=12.7 Hz), 146.7 (d, J=12.8 Hz), 142.8,140.7, 139.0, 138.0 (d, J=12.1 Hz), 130.2, 130.0, 129.4, 129.0, 128.9,128.4, 120.9, 118.8 (d, J=18.3 Hz), 118.3-117.5 (m), 116.7, 110.8 (d,J=22.2 Hz), 45.3, 44.3, 38.8. ¹⁹F NMR (377 MHz, Acetone-d₆) δ−138.7-−138.8 (m), −146.1-−146.4 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₂₁H₁₈F₂N₃O₃: 398.1316, found: 398.1312.

4-(2-((cyanomethyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide(30)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol), in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and 2-aminoacetonitrilehydrochloride (36 mg, 0.4 mmol) at room temperature. The mixture wasstirred for 2 h at room temperature and poured into a saturated solutionof ammonium chloride. After extraction with CH₂Cl₂ (3×20 mL), thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The resulting residue was purified by flash chromatography(Hexanes:EtOAc=7:3 v/v) to afford4-(2-((cyanomethyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide30 as a brownish powder (30%, 34 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.43 (s, 1H), 8.07 (s, 1H), 8.06-7.71 (m, 1H), 7.54-7.40(m, 1H), 7.37-7.12 (m, 1H), 6.93 (s, 1H), 6.88-6.84 (m, 1H), 3.93 (s,3H), 3.66-3.43 (m, 1H), 1.99-1.93 (m, 3H), 1.79-1.68 (m, 2H), 1.67-1.55(m, 2H). ¹³C NMR (101 MHz, Acetone-d₆) δ 181.1, 164.1, 161.1, 139.0,129.3, 120.5, 118.9 (d, J=17.6 Hz), 118.0, 117.8 (dd, J=6.0, 3.2 Hz),116.6, 110.9 (d, J=22.1 Hz), 38.8, 28.8. ¹⁹F NMR (377 MHz, Acetone-d₆) δ−138.7-−138.8 (m), −145.9-−146.2 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₁₇H₂₀F₂N₃O₃S: 384.1193, found: 384.1186.

N-(3,4-difluorophenyl)-1-methyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxamide(31)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol), in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and morpholine (34 μL, 0.4mmol) at room temperature. The mixture was stirred for 2 h at roomtemperature and poured into a saturated solution of ammonium chloride.After extraction with CH₂Cl₂ (3×20 mL), the combined organic layers weredried over sodium sulfate and concentrated in vacuo. The resultingresidue was purified by flash chromatography (Hexanes:EtOAc=7:3 v/v) toaffordN-(3,4-difluorophenyl)-1-methyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxamide31 as a white powder (80%, 97 mg, 0.2 mmol). ¹H NMR (400 MHz, DMSO-d₆) δ10.28 (s, 1H), 8.19-7.80 (m, 2H), 7.57-7.47 (m, 2H), 7.46-7.36 (m, 1H),3.95 (s, 3H), 3.81-3.67 (m, 2H), 3.64-3.59 (m, 2H), 3.58-3.49 (m, 2H),3.34-3.25 (m, 2H). ¹³C NMR (101 MHz, DMSO-d₆) δ 185.9, 165.8, 159.5,150.5 (d, J=13.1 Hz), 148.1 (d, J=13.2 Hz), 147.1 (d, J=12.8 Hz), 144.7(d, J=12.8 Hz), 136.4 (dd, J=9.1, 3.0 Hz), 135.9, 128.1, 119.8, 117.8(d, J=17.7 Hz), 117.2-116.3 (m), 114.0, 109.5 (d, J=21.7 Hz), 66.5 (d,J=30.5 Hz), 46.3, 41.5, 37.8. ¹⁹F NMR (377 MHz, DMSO-d₆) δ −137.2-−137.4(m), −144.5-−144.6 (m). HRMS (ESI): m/z [M+H]⁺ calcd for C₁₈H₁₈F₂N₃O₄:378.1265, found: 378.1257.

4-(2-(3,3-difluoroazetidin-1-yl)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide(32)

To a solution of2-(5-((3,4-difluorophenyl)carbamoyl)-1-methyl-1H-pyrrol-3-yl)-2-oxoaceticacid 24 (0.1 g, 0.3 mmol), in DMF (4 mL) and CH₂Cl₂ (5 mL) were added1,1′-carbonyldiimidazole (79 mg, 0.05 mmol) and 3,3-difluoroazetidinehydrochloride (50 mg, 0.4 mmol) at room temperature. The mixture wasstirred for 2 h at room temperature and poured into a saturated solutionof ammonium chloride. After extraction with CH₂Cl₂ (3×20 mL), thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The resulting residue was purified by flash chromatography(Hexanes:EtOAc=7:3 v/v) to afford4-(2-(3,3-difluoroazetidin-1-yl)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide32 as a white powder (48%, 60 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.60 (s, 1H), 8.06 (d, J=1.7 Hz, 1H), 8.01-7.86 (m, 1H),7.70-7.47 (m, 2H), 7.34-7.26 (m, 1H), 4.98-4.79 (m, 2H), 4.65-4.31 (m,2H), 4.05 (s, 3H). ¹³C NMR (101 MHz, Acetone-d₆) δ 182.6, 163.7, 161.1,152.6 (d, J=13.1 Hz), 150.2 (d, J=13.6 Hz), 149.1 (d, J=12.8 Hz), 146.7(d, J=13.1 Hz), 138.4, 138.2-137.6 (m), 129.2, 121.4, 121.0, 118.9 (d,J=17.9 Hz), 118.3, 118.0-117.5 (m), 116.2, 115.6, 110.8 (d, J=22.2 Hz),65.8 (t, J=28.6 Hz), 61.9 (t, J=28.7 Hz), 38.8. ¹⁹F NMR (377 MHz,Acetone-d₆) δ −102.1 (p, J=12.3 Hz), −138.7-−138.8 (m), −146.0-−146.2(m). HRMS (ESI): m/z [M+H]⁺ calcd for C₁₇H₁₄F₄N₃O₃: 384.0971, found:384.0964.

N-(3,4-difluorophenyl)-1-methyl-4-(phenylsulfonamido)-1H-pyrrole-2-carboxamide(36)

To a solution of 1-methyl-1H-pyrrole-2-carboxylic acid 14 (4 g, 32 mmol)in acetic anhydride (40 mL) was added nitric acid 70% (3.2 mL) at −25°C. The reaction was allowed to warm up to room temperature and wasstirred for 2 h. After addition of water (200 mL) at −25° C., themixture was extracted with ethyl acetate (3×50 mL).

The combined organic layers were washed with a saturated solution ofsodium carbonate (3×100 mL), dried over sodium sulfate, and concentratedin vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=4:6 v/v) to give compound 33 as a dark oil (14%, 770 mg,4.5 mmol). To a solution of 1-methyl-4-nitro-1H-pyrrole-2-carboxylicacid 33 (0.77 g, 4.5 mmol), 3,4-difluoroaniline (1.16 g, 9.0 mmol) andDIPEA (1.85 mL, 13.6 mmol) in DMF (20 mL) was added HATU (1.89 g, 5.0mmol) at room temperature. The reaction mixture was stirred at 50° C.overnight and then poured into a saturated solution of ammonium chloride(50 mL). After extraction with ethyl acetate (3×50 mL), the combinedorganic layers were dried over sodium sulfate and concentrated in vacuo.The residue was purified by flash chromatography (Hexanes:EtOAc=7:3 v/v)to give N-(3,4-difluorophenyl)-1-methyl-4-nitro-1H-pyrrole-2-carboxamide34 (64%, 820 mg, 2.9 mmol). To a suspension ofN-(3,4-difluorophenyl)-1-methyl-4-nitro-1H-pyrrole-2-carboxamide 34 (250mg, 0.9 mmol) in methanol (20 mL) and formic acid (0.5 mL, 13.3 mmol)was added Zn dust (250 mg, 3.8 mmol). The mixture was stirred for 10minutes at room temperature and filtered on Celite. The solution wasextracted with HCl 1N (3×50 mL) and washed with ethyl acetate (2×50 mL).The aqueous layer was then basified using a 5% solution of sodiumhydroxide (pH>8) and extracted with ethyl acetate (3×50 mL).

The combined organic layers were dried over sodium sulfate andconcentrated in vacuo to give4-amino-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide 35 as abrown/yellowish solid (68%, 153 mg, 0.6 mmol). To a solution of4-amino-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide 35 (25mg, 0.1 mmol) in DMF (3 mL) were added benzenesulfonylchloride (14 μL,0.1 mmol) and Et₃N (20 μL, 0.2 mmol). The reaction mixture was stirredovernight at room temperature and then poured into a saturated solutionof ammonium chloride (20 mL). After extraction with EtOAc (3×20 mL), thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=1:1 v/v) to give compound 36 (51%, 20 mg, 0.05 mmol). ¹HNMR (400 MHz, Acetone-d₆) δ 9.37 (s, 1H), 8.53 (s, 1H), 8.05-7.85 (m,1H), 7.81-7.71 (m, 2H), 7.68-7.58 (m, 1H), 7.58-7.51 (m, 2H), 7.45-7.41(m, 1H), 7.35-7.17 (m, 1H), 6.77 (d, J=2.0 Hz, 1H), 6.72 (d, J=2.0 Hz,1H), 3.86 (s, 3H). ¹³C NMR (101 MHz, Acetone-d₆) δ 161.3, 141.9, 134.3,130.6, 128.8, 125.6, 124.2, 121.9, 118.7 (d, J=17.9 Hz), 118.1-117.4(m), 110.7 (d, 0.1=22.1 Hz), 110.2, 38.0. ¹⁹F NMR (377 MHz, Acetone-d₆)δ −140.2-−140.4 (m), −147.9-−148.1 (m). HRMS (ESI): m/z [M+H]⁺ calcd forC₁₈H₁₆F₂N₃O₃S: 392.0880, found: 392.0872.

4-(cyclopentanesulfonamido)-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide(37)

To a solution of4-amino-N-(3,4-difluorophenyl)-1-methyl-1H-pyrrole-2-carboxamide 35 (50mg, 0.2 mmol) in DMF (5 mL) were added cyclopentylsulfonylchloride (51μL, 0.4 mmol) and Et₃N (40 μL, 0.3 mmol). The reaction mixture wasstirred overnight at room temperature and then poured into a saturatedsolution of ammonium chloride (30 mL). After extraction with EtOAc (3×30mL), the combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=1:1 v/v) to give compound 37 (56%, 43 mg, 0.1 mmol). ¹HNMR (400 MHz, Acetone-d₆) δ 9.43 (s, 1H), 8.07 (s, 1H), 8.06-7.71 (m,1H), 7.54-7.40 (m, 1H), 7.37-7.12 (m, 1H), 6.93 (s, 1H), 6.88-6.84 (m,1H), 3.93 (s, 3H), 3.66-3.43 (m, 1H), 1.99-1.93 (m, 3H), 1.79-1.68 (m,2H), 1.67-1.55 (m, 2H). ¹⁹F NMR (377 MHz, Acetone-d₆) δ −140.2-−140.4(m), −148.0-−148.1 (m). HRMS (ESI): m/z [M+H]⁺ calcd for C₁₇H₂₀F₂N₃O₃S:384.1193, found: 384.1186.

4-(2-(((1H-1,2,3-triazol-4-yl)methyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(38)

To a solution of 7a in an ACN/H₂O mixture (2 mL, 1:1) were added sodiumazide (26 mg, 0.4 mmol), CuSO₄.5H₂O (5 mg) and sodium ascorbate (12 mg).The reaction mixture was heated at 150° C. under microwave irradiationfor 1 h. The solution was diluted with EtOAc and washed with water. Theorganic layer was dried over MgSO₄ and concentrated in vacuo. Theresidue was purified by silica gel column chromatography using DCM/MeOH(95:5) to afford 38 in 41% yield (23 mg). HRMS (ESI): m/z [M+H]⁺ calcdfor C₁₉H₁₉F₂N₆O₃: 417.1487, found: 417.1476.

1,3,5-Trimethyl-1H-pyrrole-2-carboxylic acid (39)

To a solution of 2 (3 g, 72 mmol) in EtOH (100 mL) was added NaOH 20%(70 mL). The reaction was heated at 100° C. for 6 h. EtOH was evaporatedunder vacuum and the mixture was washed with DCM (3×30 mL). The aqueouslayer was carefully acidified to pH 3-4 with 1M HCl. The mixture wasextracted with DCM (3×30 mL). Combined organic layers were dried overMgSO₄ and concentrated in vacuo. The resulting solid was washed withcold Et₂O to afford 39 in 61% yield (6.7 g) as a pink solid. ¹H NMR (400MHz, DMSO-d₆) δ 11.88 (s, 1H), 5.75 (s, 1H), 3.68 (s, 3H), 2.19 (s, 3H),2.15 (s, 3H). MS (ESI): m/z [M+H]+ calcd for C₈H₁₂NO₂: 154.1, found:154.5.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide (40)

To a solution of 39 (2.9 g, 18.9 mmol) in DMF (20 mL) were added3,4-difluoroaniline (4.5 mL, 22.7 mmol), HATU (8.6 g, 22.7 mmol) andDIPEA (6.6 mL, 37.8 mmol) at 0° C. The mixture was heated at 60° C. for2 days. The reaction mixture was then diluted with EtOAC and washed with1M HCl, water and brine. The organic layers was dried over MgSO₄ andconcentrated in vacuo. The residue was purified by silica gel columnchromatography using hexanes/EtOAc (8:2) to afford 40 in 37% yield (1.85g). ¹H NMR (400 MHz, Chloroform-d) δ 7.76-7.65 (m, 1H), 7.19-7.07 (m,2H), 5.80 (s, 1H), 3.77 (s, 3H), 2.38 (s, 3H), 2.24 (s, 3H). MS (ESI):m/z [M+H]+ calcd for C₁₄H₁₄F₂N₂O: 264.1, found: 265.5.

2-(5-((3,4-Difluorophenyl)carbamoyl)-1,2,4-trimethyl-1H-pyrrol-3-yl)-2-oxoaceticacid (42)

To a solution of 40 (860 mg, 3.26 mmol) in DCM (30 mL) were added ethyloxalylchloride (980 μL, 8.80 mmol) and AlCl₃ (1.08 g, 8.15 mmol) at 0°C. The mixture was stirred at room temperature for 16 h and poured intocrushed ice. The mixture was extracted with DCM and combined organiclayers were filtered on Celite. The filtrate was concentrated and theresulting residue was used in the next step without furtherpurification. To a solution of crude 41 in EtOH was added NaOH 10% (25mL). The mixture was stirred for 1 h at room temperature. EtOH wasevaporated under vacuum and the mixture was extracted with EtOAc (3×10mL). The aqueous layer was acidified with 1M HCl. The mixture wasextracted with EtOAc (3×10 mL). Combined organic layers were dried overMgSO₄ and concentrated in vacuo. The resulting solid was washed withEt₂O to afford 42 in 59% yield (646 mg) over two steps. ¹H NMR (400 MHz,DMSO-d₆) δ 14.13 (s, 1H), 10.46 (s, 1H), 8.04-7.71 (m, 1H), 7.59-7.28(m, 2H), 3.60 (s, 3H), 2.46 (s, 3H), 2.26 (s, 3H). MS (ESI): m/z [M+H]+calcd for C₁₆H₁₅F₂N₂O₄: 337.1, found: 337.5.

General Procedure for the Synthesis of 43-48

To a solution of 42 (40 mg, 0.119 mmol) in a DMF/DCM mixture (2 mL, 1:1)was added CDI (29 mg, 0.178 mmol) at room temperature. After 15 min, theamine (0.178 mmol) was added and the mixture was stirred for 1 h. Thereaction mixture was diluted with EtOAc and washed with H₂O (3×5 mL).The organic layer was dried over MgSO₄ and concentrated in vacuo. Theresidue was purified by silica gel column chromatography using DCM/MeOH(98:2) to afford compounds 43-48.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(pyridin-2-ylamino)acetyl)-1H-pyrrole-2-carboxamide(43)

Yield: 69%. ¹H NMR (400 MHz, Chloroform-d) δ 9.33 (s, 1H), 8.45-8.38 (m,1H), 8.30 (d, J=8.3 Hz, 1H), 7.85-7.67 (m, 2H), 7.49 (s, 1H), 7.22-7.12(m, 3H), 3.75 (s, 3H), 2.45 (s, 3H), 2.44 (s, 3H). FIRMS (ESI): m/z[M+H]⁺ calcd for C₂₁H₁₉F₂N₄O₃: 413.1425, found: 413.1416.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-(4-methylpiperazin-1-yl)-2-oxoacetyl)-1H-pyrrole-2-carboxamide(44)

Yield: 74%. ¹H NMR (400 MHz, Chloroform-d) δ 8.98 (s, 1H), 7.80-7.69 (m,1H), 7.43-7.32 (m, 1H), 7.13 (dt, J=10.0, 8.8 Hz, 1H), 3.72-3.67 (m,2H), 3.66 (s, 3H), 3.32 (dd, J=5.9, 4.1 Hz, 2H), 2.46 (t, J=5.2 Hz, 2H),2.42 (s, 3H), 2.35 (t, J=5.1 Hz, 2H), 2.31 (s, 3H), 2.30 (s, 3H). HRMS(ESI): m/z [M+H]⁺ calcd for C₂₁H₂₅F₂N₄O₃: 419.1895, found: 419.1886.

4-(2-(diethylamino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(45)

Yield: 71%. ¹H NMR (400 MHz, Chloroform-d) δ 9.07 (s, 1H), 7.87-7.73 (m,1H), 7.45-7.36 (m, 1H), 7.13 (dt, J=10.0, 8.8 Hz, 1H), 3.66 (s, 3H),3.48 (q, J=7.1 Hz, 2H), 3.22 (q, J=7.0 Hz, 2H), 2.42 (s, 3H), 2.31 (s,3H), 1.21 (t, J=7.1 Hz, 3H), 1.14 (t, J=7.0 Hz, 3H). HRMS (ESI): m/z[M+H]⁺ calcd for C₂₀H₂₄F₂N₃O₃: 392.1786, found: 392.1776.

4-(2-((1H-benzo[d]imidazol-2-yl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(46)

Yield: 46%. ¹H NMR (400 MHz, DMSO-d₆) δ 12.42 (s, 2H), 10.45 (s, 1H),8.03-7.76 (m, 1H), 7.56-7.38 (m, 4H), 7.18 (dd, J=5.9, 3.2 Hz, 2H), 3.61(s, 3H), 2.45 (s, 3H), 2.27 (s, 3H). FIRMS (ESI): m/z [M+H]⁺ calcd forC₂₃H₂₀F₂N₅O₃: 452.1534, found: 452.1525.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-((pyridin-2-ylmethyl)amino)acetyl)-1H-pyrrole-2-carboxamide(47)

Yield: 55%. ¹H NMR (400 MHz, DMSO-d₆) δ 10.42 (s, 1H), 9.28 (t, J=6.1Hz, 1H), 8.53 (d, J=4.8 Hz, 1H), 7.94-7.77 (m, 2H), 7.46-7.41 (m, 2H),7.38 (d, J=8.0 Hz, 1H), 7.34-7.26 (m, 1H), 4.50 (d, J=5.9 Hz, 2H), 3.59(s, 3H), 2.38 (s, 3H), 2.21 (s, 3H). HRMS (EST): m/z [M+H]⁺ calcd forC₂₂H₂₁F₂N₄O₃: 427.1582, found: 427.1572.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-(((1-methyl-1H-imidazol-2-yl)methyl)amino)-2-oxoacetyl)-1H-pyrrole-2-carboxamide(48)

Yield: 63%. ¹H NMR (400 MHz, Chloroform-d) δ 8.48 (s, 2H), 7.77-7.66 (m,1H), 7.26-7.20 (m, 1H), 7.13 (dt, J=9.9, 8.7 Hz, 1H), 6.93 (d, J=1.3 Hz,1H), 6.85 (d, J=1.3 Hz, 1H), 4.53 (d, =5.7 Hz, 2H), 3.72 (s, 3H), 3.64(s, 3H), 2.30 (s, 3H), 2.24 (s, 3H). HRMS (ESI): m/z [M+H]⁺ calcd forC₂₁H₂₂F₂N₅O₃: 430.1691, found: 430.1681.

1,3,5-trimethyl-N-phenyl-1H-pyrrole-2-carboxamide (49)

To a solution of 39 (2.0 g, 13 mmol) in DMF (50 mL) were added aniline(2.4 mL, 26 mmol), HATU (5.93 g, 15.6 mmol) and DIPEA (4.5 mL, 26 mmol)at 0° C. The mixture was heated at 60° C. for 2 days. The reactionmixture was then diluted with EtOAc and washed with 1M HCl, water andbrine. The organic layers was dried over MgSO₄ and concentrated invacuo. The residue was purified by silica gel column chromatographyusing hexanes/EtOAc (8:2) to afford 49 in 58% yield (1.72 g). ¹H NMR(400 MHz, DMSO-d₆) δ 9.60 (s, 1H), 7.68 (d, J=8.0 Hz, 2H), 7.31 (t,J=7.7 Hz, 2H), 7.04 (t, J=7.4 Hz, 1H), 5.73 (s, 1H), 3.57 (s, 3H), 2.17(s, 6H). MS (ESI): m/z [M+H]+ calcd for C₁₄H₁₇N₂O: 229.1, found: 229.5.

2-oxo-2-(1,2,4-trimethyl-5-(phenylcarbamoyl)-1H-pyrrol-3-yl)acetic acid(51)

To a solution of 49 (695 mg, 3.05 mmol) in DCM (30 mL) were added ethyloxalylchloride (916 μL, 8.23 mmol) and AlCl₃ (1.01 g, 7.62 mmol) at 0°C. The mixture was stirred at room temperature for 16 h and poured intocrushed ice. The mixture was extracted with DCM and combined organiclayers were filtered on Celite. The filtrate was concentrated and theresulting residue was used in the next step without furtherpurification. To a solution of crude 50 in EtOH was added NaOH 10% (30mL). The mixture was stirred for 1 h at room temperature. EtOH wasevaporated under vacuum and the mixture was washed with EtOAc (3×10 mL).The aqueous layer was acidified with 1M HCl. The mixture was extractedwith EtOAc (3×10 mL). Combined organic layers were dried over MgSO₄ andconcentrated in vacuo. The resulting solid was washed with Et₂O toafford 51 in 70% yield (642 mg) over two steps. ¹H NMR (400 MHz,DMSO-d₆) δ 14.18 (s, 1H), 10.26 (s, 1H), 7.71 (d, 0.1=8.0 Hz, 2H), 7.35(t, 0.1=7.8 Hz, 2H), 7.10 (td, 0.1=7.4, 1.1 Hz, 1H), 3.61 (s, 3H), 2.46(s, 3H), 2.26 (s, 3H). MS (ESI): m/z [M+H]+ calcd for C₁₆H₁₇N₂O₄: 301.1,found: 301.5.

General Procedure for the Synthesis of Compounds 52 and 53

To a solution of 51 (50 mg, 0.17 mmol) in DMF (2 mL) were added amine(0.25 mmol), HATU (76 mg, 0.20 mmol) and DIPEA (58 μL, 0.33 mmol) at 0°C. The mixture was stirred at room temperature for 2 h. The solution wasthen diluted with EtOAc and washed with water and brine. The organiclayers was dried over MgSO₄ and concentrated in vacuo. The residue waspurified by silica gel column chromatography using DCM/MeOH (98:2) toafford to afford the desired compounds 52 and 53.

1,3,5-trimethyl-4-(2-oxo-2-(pyridin-2-ylamino)acetyl)-N-phenyl-1H-pyrrole-2-carboxamide(52)

Yield: 75%. ¹H NMR (400 MHz, Chloroform-d) δ 9.47 (s, 1H), 8.43-8.35 (m,1H), 8.30 (d, J=8.3 Hz, 1H), 7.86-7.74 (m, 1H), 7.68-7.57 (m, 3H),7.46-7.34 (m, 1H), 7.22-7.10 (m, 2H), 3.73 (s, 3H), 2.44 (s, 3H), 2.43(s, 3H). MS (ESI): m/z [M+H]+ calcd for C₂₁H₂₁N₄O₃: 377.2, found: 377.4.

4-(2-((5-fluoropyridin-2-yl)amino)-2-oxoacetyl)-1,3,5-trimethyl-N-phenyl-1H-pyrrole-2-carboxamide(53)

Yield: 68%. ¹H NMR (400 MHz, DMSO-d₆) δ 11.16 (s, 1H), 10.26 (s, 1H),8.61-8.47 (m, 1H), 8.40-8.19 (m, 1H), 7.76-7.66 (m, 2H), 7.34 (dd,J=8.5, 7.4 Hz, 2H), 7.25 (dd, J=8.8, 3.2 Hz, 1H), 7.15-7.04 (m, 1H),3.62 (s, 3H), 2.44 (s, 3H), 2.26 (s, 3H). MS (ESI): m/z [M+H]+ calcd forC₂₁H₂₀FN₄O₃: 395.2, found: 395.5.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(4-(trifluoromethoxy)phenyl)-1H-pyrrole-2-carboxamide(54)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (250 mg, 1.0 mmol), 4-(trifluoromethoxy)aniline (227 mg, 1.3mmol) and DIPEA (330 μL, 2.0 mmol) in pyridine (10 mL) was added HATU(0.65 g, 1.8 mmol) at room temperature. The mixture was stirred at 65°C. for 18 h. The reaction mixture was then poured into a saturatedsolution of ammonium chloride and extracted with ethyl acetate (3×50mL). The combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 54 as a white powder (35%, 141mg, 0.3 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.51 (s, 1H), 8.17 (s,1H), 7.95 (d, 0.1=9.1 Hz, 2H), 7.36 (d, 0.1=8.1 Hz, 2H), 4.17 (dd,0.1=5.9, 2.6 Hz, 2H), 3.70 (s, 3H), 2.75 (t, J=2.5 Hz, 1H), 2.45 (s,3H), 2.31 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd for C₂₀H₁₉F₃N₃O₄: 422.4,found: 422.4.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(3-(trifluoromethoxy)phenyl)-1H-pyrrole-2-carboxamide(55)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (250 mg, 1.0 mmol), 3-(trifluoromethoxy)aniline (227 mg, 1.3mmol) and DIPEA (330 μL, 2.0 mmol) in pyridine (10 mL) was added HATU(0.65 g, 1.8 mmol) at room temperature. The mixture was stirred at 65°C. for 18 h. The reaction mixture was then poured into a saturatedsolution of ammonium chloride and extracted with ethyl acetate (3×50mL). The combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 55 as a white powder (34%, 135mg, 0.3 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.57 (s, 1H), 8.18 (s,1H), 8.02 (s, 1H), 7.81-7.67 (m, 1H), 7.50 (t, J=8.2 Hz, 1H), 7.16-7.06(m, 1H), 4.17 (dd, J=5.8, 2.6 Hz, 2H), 3.71 (s, 3H), 2.75 (t, J=2.5 Hz,1H), 2.45 (s, 3H), 2.32 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₂₀H₁₉F₃N₃O₄: 422.4, found: 422.4.

N-(2-(3,4-difluorophenyl)propan-2-yl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(56)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol), 2-(3,4-difluorophenyl)propan-2-amine (65 mg,0.4 mmol) and DIPEA (132 μL, 0.8 mmol) in DMF (5 mL) was added HATU(0.218 g, 0.6 mmol) at room temperature. The mixture was stirred at 65°C. for 18 h. The reaction mixture was then poured into a saturatedsolution of ammonium chloride and extracted with ethyl acetate (3×50mL). The combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 56 (38%, 61 mg, 0.1 mmol). ¹HNMR (400 MHz, Acetone-d₆) δ 8.11 (s, 1H), 7.59 (s, 1H), 7.47-7.38 (m,1H), 7.37-7.31 (m, 1H), 7.26 (dt, =10.5, 8.5 Hz, 1H), 4.14 (dd, =5.8,2.5 Hz, 2H), 3.53 (s, 3H), 2.72 (t, J=2.6 Hz, 1H), 2.37 (s, 3H), 2.30(s, 3H), 1.77 (s, 6H). MS (ESI): m/z [M+H]⁺ calcd for C₂₂H₂₄F₂N₃O₃:416.4, found: 416.5.

N-(1-(3,4-difluorophenyl)cyclopropyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(57)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (75 mg, 0.3 mmol), 1-(3,4-difluorophenyl)cyclopropanamine (50 mg,0.3 mmol) and DIPEA (100 μL, 0.6 mmol) in DMF (5 mL) was added HATU(0.163 g, 0.4 mmol) at room temperature. The mixture was stirred at 65°C. for 18 h. The reaction mixture was then poured into a saturatedsolution of ammonium chloride and extracted with ethyl acetate (3×50mL). The combined organic layers were dried over sodium sulfate andconcentrated in vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 57 (46%, 55 mg, 0.1 mmol). ¹HNMR (400 MHz, Acetone-d₆) δ 8.11 (s, 1H), 8.06 (s, 1H), 7.35-7.28 (m,1H), 7.27-7.17 (m, 2H), 4.13 (dd, J=5.8, 2.5 Hz, 2H), 3.60 (s, 3H), 2.72(t, J=2.6 Hz, 1H), 2.38 (s, 3H), 2.23 (s, 3H), 1.41-1.30 (m, 4H). MS(ESI): m/z [M+H]⁺ calcd for C₂₂H₂₂F₂N₃O₃: 414.4, found: 414.5.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-phenyl-1H-pyrrole-2-carboxamide(58)

To a solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol), aniline (53 mg, 0.6 mmol) and DIPEA (100 μL,0.6 mmol) in DMF (5 mL) was added HATU (0.163 g, 0.4 mmol) at roomtemperature. The mixture was stirred at 65° C. for 18 h. The reactionmixture was then poured into a saturated solution of ammonium chlorideand extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (DCM:MeOH=98:2 v/v) to givecompound 58 as a yellowish powder (66%, 82 mg, 0.2 mmol). ¹H NMR (400MHz, Acetone-d₆) δ 9.32 (s, 1H), 8.18 (s, 1H), 7.88-7.79 (m, 2H),7.44-7.32 (m, 1H), 7.20-7.08 (m, 1H), 4.16 (dd, J=5.9, 2.5 Hz, 2H), 3.69(s, 3H), 2.75 (t, J=2.5 Hz, 1H), 2.44 (s, 3H), 2.31 (s, 3H). MS (ESI):m/z [M+H]⁺ calcd for C₁₉H₂₀N₃O₃: 338.4, found: 338.5.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(4-(pentafluorosulfanyl)phenyl)-1H-pyrrole-2-carboxamide(59)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 4-pentafluorosulfanylaniline (167 mg, 0.8 mmol)in N,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0°C. for 2 h, poured into a solution of hydrochloric acid 1N (50 mL), andextracted with ethyl acetate (3×50 mL). The combined organic layers weredried over sodium sulfate and concentrated in vacuo. The residue waspurified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 59 as a white powder (47%, 83 mg, 0.2 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.72 (s, 1H), 8.19 (s, 1H), 8.03 (d, J=8.8 Hz, 2H), 7.88(d, J=9.3 Hz, 2H), 4.16 (dd, J=5.9, 2.6 Hz, 2H), 3.71 (s, 3H), 2.75 (t,J=2.5 Hz, 1H), 2.45 (s, 3H), 2.32 (s, 3H). MS (ESI): m/z [M+H]⁺ calcdfor C₂₀H₁₉F₃N₃O₄: 464.4, found: 464.4.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(3-(pentafluorosulfanyl)phenyl)-1H-pyrrole-2-carboxamide(60)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 3-pentafluorosulfanylaniline (167 mg, 0.8 mmol)in N,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0°C. for 2 h, poured into a solution of hydrochloric acid 1N (50 mL), andextracted with ethyl acetate (3×50 mL). The combined organic layers weredried over sodium sulfate and concentrated in vacuo. The residue waspurified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 60 as a white powder (40%, 71 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.68 (s, 1H), 8.54 (s, 1H), 8.20 (s, 1H), 8.00 (m, 1H),7.63 (m, 1H), 4.16 (dd, J=5.9, 2.6 Hz, 2H), 3.71 (s, 3H), 2.75 (t, J=2.5Hz, 1H), 2.45 (s, 3H), 2.33 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₁₉H₁₉F₅N₃O₃S: 464.4, found: 464.4.

N-(6-fluoropyridin-3-yl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(61)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 6-fluoropyridin-3-amine (85 mg, 0.8 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 61 as a white powder (68%, 92 mg, 0.2 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.56 (s, 1H), 8.62 (s, 1H), 8.47-8.36 (m, 1H), 8.18 (s,1H), 7.14 (dd, 0.1=8.9, 3.4 Hz, 1H), 4.24-4.15 (m, 2H), 3.72 (s, 3H),2.78-2.74 (m, 1H), 2.46 (s, 3H), 2.34 (s, 3H). MS (ESI): m/z [M+H]⁺calcd for C₁₈H₁₈FN₄O₃: 357.4, found: 357.4.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(pyrimidin-5-yl)-1H-pyrrole-2-carboxamide(62)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of pyrimidin-5-amine (73 mg, 0.8 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 62 as a white powder (31%, 40 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.61 (s, 1H), 9.21 (s, 2H), 8.93 (s, 1H), 8.19 (s, 1H),4.19 (dd, J=5.8, 2.6 Hz, 2H), 3.75 (s, 3H), 2.77 (t, J=2.6 Hz, 1H), 2.47(s, 3H), 2.37 (s, 3H). MS (ESI): m/z [M+h]⁺ calcd for C₁₇H₁₈N₅O₃: 340.4,found: 340.5.

1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-N-(pyridin-4-yl)-1H-pyrrole-2-carboxamide(63)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 4-aminopyridine (53 mg, 0.6 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (DCM:MeOH=98:2 v/v) to givecompound 63 (71%, 92 mg, 0.3 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.60(s, 1H), 8.49 (d, 0.1=6.5 Hz, 2H), 8.19 (s, 1H), 7.75 (d, 0.1=6.5 Hz,2H), 4.15 (dd, 0.1=5.7, 2.5 Hz, 2H), 3.70 (s, 3H), 2.74 (t, J=2.6 Hz,1H), 2.44 (s, 3H), 2.30 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₁₈H₁₉N₄O₃: 339.4, found: 339.5.

N-(4-fluoro-3-methylphenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(64)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μl, 2.7 mmol) intoluene (5 mL) was refluxed for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 4-fluoro-3-methylaniline (71 mg, 0.6 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 64 as a white solid (67%, 95 mg, 0.2 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.25 (s, 1H), 8.14 (s, 1H), 7.72 (dd, J=6.9, 2.5 Hz, 1H),7.68-7.59 (m, 1H), 7.05 (t, J=9.2 Hz, 1H), 4.15 (dd, J=5.9, 2.6 Hz, 2H),3.68 (s, 3H), 2.73 (t, J=2.6 Hz, 1H), 2.42 (s, 3H), 2.28 (s, 3H), 2.27(d, J=2.0 Hz, 3H). MS (ESI): m/z [M+H]⁺ calcd for C₂₀H₂₁FN₃O₃: 370.4,found: 370.5.

N-(3-cyano-4-fluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(65)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was refluxed for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 5-amino-2-fluorobenzonitrile (78 mg, 0.6 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 65 as a white solid (42%, 61 mg, 0.2 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.61 (s, 1H), 8.32 (dd, J=5.7, 2.7 Hz, 1H), 8.16 (s, 1H),8.13-8.07 (m, 1H), 7.46 (t, J=9.1 Hz, 1H), 4.15 (dd, J=5.8, 2.5 Hz, 2H),3.70 (s, 3H), 2.74 (t, J=2.5 Hz, 1H), 2.44 (s, 3H), 2.30 (s, 3H). MS(ESI): m/z [M+H]⁺ calcd for C₂₀H₁₈FN₄O₃: 381.4, found: 381.3.

N-(3-chloro-4-fluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(prop-2-yn-1-ylamino)acetyl)-1H-pyrrole-2-carboxamide(66)

A solution of4-[2-(propargylamino)-2-oxo-acetyl]-1,3,5-trimethyl-pyrrol-2-carboxylicacid 6 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was refluxed for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 3-chloro-4-fluoroaniline (83 mg, 0.6 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 66 as a white solid (20%, 30 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.45 (s, 1H), 8.29-8.03 (m, 2H), 7.84-7.64 (m, 1H), 7.32(t, J=9.0 Hz, 1H), 4.15 (dd, J=5.9, 2.6 Hz, 2H), 3.69 (s, 3H), 2.73 (s,1H), 2.43 (s, 3H), 2.29 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₁₉H₁₈ClFN₃O₃: 390.8, found: 390.4.

1,3,5-trimethyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxylicacid (68)

To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(2.5 g, 9.9 mmol) in DMF (15 mL) and CH₂Cl₂ (10 mL) was added1,1′-carbonyldiimidazole (2.4 g, 11.8 mmol) and aniline (1.35 mL, 9.5mmol). After stirring for 2 h at room temperature, the reaction mixturewas poured onto a saturated solution of ammonium chloride and extractedwith CH₂Cl₂ (3×100 mL). The combined organic layers were dried oversodium sulfate and concentrated in vacuo to give 67 as a white solid. Tothe crude ethyl1,3,5-trimethyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxylate67 dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solutionof sodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted again with ethylacetate (3×50 mL). The combined organic layers were dried over sodiumsulfate and concentrated in vacuo. The resulting solid was washed withdiethyl ether (50 mL) and hexanes (50 mL) to yield1,3,5-trimethyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxylicacid 68 (1.8 g, 6.0 mmol, 62%). ¹H NMR (400 MHz, DMSO-d₆) δ 10.79 (s,1H), 7.75-7.67 (m, 2H), 7.37 (t, J=7.9 Hz, 2H), 7.20-7.11 (m, 1H), 3.77(s, 3H), 2.41 (s, 6H). MS (ESI): m/z [M+H]⁺ calcd for C₁₆H₁₇N₂O₄: 301.3,found: 301.4.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxamide(69)

To a solution of1,3,5-trimethyl-4-(2-oxo-2-(phenylamino)acetyl)-1H-pyrrole-2-carboxylicacid 68 (200 mg, 0.7 mmol), 3,4-difluoroaniline (129 mg, 1.0 mmol) andDIPEA (231 μL, 4.3 mmol) in DMF (5 mL) was added HATU (304 mg, 0.8 mmol)at room temperature. The mixture was stirred at 50° C. for 3 h. In orderto reach completion, more 3,4-difluoroaniline (65 mg, 0.5 mmol) wasadded and the mixture was further stirred overnight at 65° C. Thereaction mixture was then poured into a saturated solution of ammoniumchloride and extracted with ethyl acetate (3×50 mL). The combinedorganic layers were dried over sodium sulfate and concentrated in vacuo.The residue was purified by flash chromatography (Hexanes:EtOAc=6:4 v/v)to give compound 69 as a white powder (35%, 110 mg, 0.3 mmol). ¹H NMR(400 MHz, Acetone-d₆) δ 9.73 (s, 1H), 9.52 (s, 1H), 8.02-7.94 (m, 1H),7.87-7.80 (m, 2H), 7.56-7.48 (m, 1H), 7.43-7.36 (m, 2H), 7.32 (dt,J=10.6, 9.0 Hz, 1H), 7.20-7.13 (m, 1H), 3.70 (s, 3H), 2.46 (s, 3H), 2.31(s, 3H). MS (ESI): m/z [M+H]⁺ calcd for C₂₂H₂₀F₂N₃O₃: 412.4, found:412.5.

1,3,5-trimethyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxylicacid (71)

To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(0.5 g, 2.0 mmol) in DMF (15 mL) and CH₂Cl₂ (10 mL) was added1,1′-carbonyldiimidazole (0.53 g, 3.2 mmol) and morpholine (0.25 mL, 3.2mmol). After stirring for 2 h at room temperature, the reaction mixturewas poured onto a saturated solution of ammonium chloride and extractedwith CH₂Cl₂ (3×100 mL). The combined organic layers were dried oversodium sulfate and concentrated in vacuo to give 70 as a yellowish oil.To the crude ethyl1,3,5-trimethyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxylate 70dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solution ofsodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted again with ethylacetate (3×50 mL). The combined organic layers were dried over sodiumsulfate and concentrated in vacuo. The resulting solid was washed withdiethyl ether (50 mL) and hexanes (50 mL) to yield1,3,5-trimethyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxylicacid 71 (0.41 g, 1.3 mmol, 70%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.86 (s,1H), 3.75 (s, 3H), 3.71-3.63 (m, 2H), 3.59-3.51 (m, 4H), 3.33-3.25 (m,2H), 2.44 (s, 3H), 2.42 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₁₄H₁₉N₂O₅: 295.3, found: 295.4.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxamide(72)

A solution of1,3,5-trimethyl-4-(2-morpholino-2-oxoacetyl)-1H-pyrrole-2-carboxylicacid 71 (100 mg, 0.3 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was heated to reflux for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 3,4-difluoroaniline (66 mg, 0.5 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (DCM:MeOH=98:2 v/v) to givecompound 72 as a brownish solid (29%, 40 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.53 (s, 1H), 8.03-7.93 (m, 1H), 7.58-7.44 (m, 1H), 7.33(dt, J=10.6, 9.0 Hz, 1H), 3.74-3.67 (m, 6H), 3.66-3.61 (m, 3H), 3.39 (t,J=4.8 Hz, 2H), 2.51 (s, 3H), 2.33 (s, 3H). MS (ESI): m/z [M+H]⁺ calcdfor C₂₀H₂₂F₂N₃O₄: 406.4, found: 406.5.

4-(2-(allylamino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid (74)

To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(0.5 g, 2.0 mmol) in DMF (15 mL) and CH₂Cl₂ (10 mL) was added1,1′-carbonyldiimidazole (0.53 g, 3.2 mmol) and allylamine (0.18, 3.2mmol). After stirring for 2 h at room temperature, the reaction mixturewas poured onto a saturated solution of ammonium chloride and extractedwith CH₂Cl₂ (3×100 mL). The combined organic layers were dried oversodium sulfate and concentrated in vacuo to give 73 as a white solid. Tothe crude ethyl4-(2-(allylamino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylate73 dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solutionof sodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted again with ethylacetate (3×50 mL). The combined organic layers were dried over sodiumsulfate and concentrated in vacuo. The resulting solid was washed withdiethyl ether (50 mL) and hexanes (50 mL) to yield4-(2-(allylamino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid 74 (270 mg, 1.0 mmol, 51%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.74 (s,1H), 8.86 (t, J=5.8 Hz, 1H), 5.99-5.66 (m, 1H), 5.36-5.05 (m, 2H),3.85-3.80 (m, 2H), 3.75 (s, 3H), 2.36 (s, 6H). MS (ESI): m/z [M+H]⁺calcd for C₁₃H₁₇N₂O₄: 265.3, found: 265.4.

4-(2-(allylamino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(75)

A solution of4-(2-(allylamino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid 74 (100 mg, 0.4 mmol) and thionyl chloride (210 μL, 2.7 mmol) intoluene (5 mL) was refluxed for 1 h. The resulting solution wasconcentrated in vacuo, solubilized in N,N-dimethylacetamide (5 mL), andadded to a solution of 3,4-difluoroaniline (73 mg, 0.6 mmol) inN,N-dimethylacetamide (5 mL) at 0° C. The mixture was stirred at 0° C.for 2 h, poured into a saturated solution of ammonium chloride (50 mL),and extracted with ethyl acetate (3×50 mL). The combined organic layerswere dried over sodium sulfate and concentrated in vacuo. The residuewas purified by flash chromatography (Hexanes:EtOAc=6:4 v/v) to givecompound 75 as a white solid (18%, 25 mg, 0.1 mmol). ¹H NMR (400 MHz,Acetone-d₆) δ 9.48 (s, 1H), 8.09-7.94 (m, 1H), 7.89 (s, 1H), 7.59-7.48(m, 1H), 7.32 (dt, J=10.2, 9.0 Hz, 1H), 6.02-5.86 (m, 1H), 5.28 (dd,J=17.2, 1.6 Hz, 1H), 5.12 (dd, J=10.2, 1.4 Hz, 1H), 3.99-3.94 (m, 2H),3.68 (s, 3H), 2.42 (s, 3H), 2.28 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd forC₁₉H₂₀F₂N₃O₃: 376.4, found: 376.4.

1,3,5-trimethyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid (77)

To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(2.0 g, 7.9 mmol) in DMF (15 mL) and CH₂Cl₂ (10 mL) was added1,1′-carbonyldiimidazole (1.92 g, 11.8 mmol) and 2-aminothiazole (0.95g, 9.5 mmol). After stirring for 2 h at room temperature, the reactionmixture was poured onto a saturated solution of ammonium chloride,filtered through fritted funnel, dried under vacuum to give 76 as ayellow solid. To the crude ethyl1,3,5-trimethyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxylate76 dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solutionof sodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted again with ethylacetate (3×50 mL). The combined organic layers were dried over sodiumsulfate and concentrated in vacuo. The resulting solid was washed withdiethyl ether (50 mL) and hexanes (50 mL) to yield1,3,5-trimethyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid 77 (1.9 g, 6.1 mmol, 78%) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ 12.96 (s, 1H), 7.58 (s, 1H), 7.38 (s, 1H), 3.77 (s, 3H), 2.37(s, 3H), 2.33 (s, 3H). MS (ESI): m/z [1\4-HE]⁺ calcd for C₁₃H₁₄N₃O₄:308.3, found: 308.4.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxamide(78)

To a solution of1,3,5-trimethyl-4-(2-oxo-2-(thiazol-2-ylamino)acetyl)-1H-pyrrole-2-carboxylicacid 77 (250 mg, 0.8 mmol), 3,4-difluoroaniline (158 mg, 1.2 mmol) andDIPEA (283 μL, 1.6 mmol) in DMF (15 mL) was added HATU (0.37 g, 1.0mmol) at room temperature. The mixture was stirred at 50° C. for 3 h. Inorder to reach completion, more 3,4-difluoroaniline (80 mg, 0.6 mmol)was added and the mixture was further stirred overnight at 65° C. Thereaction mixture was then poured into a saturated solution of ammoniumchloride and extracted with ethyl acetate (3×50 mL). The combinedorganic layers were dried over sodium sulfate and concentrated in vacuo.The residue was purified by flash chromatography (DCM:MeOH=98:2 v/v) togive compound 78 as a yellowish powder (55%, 187 mg, 0.4 mmol). ¹H NMR(400 MHz, DMSO-d₆) δ 13.01 (s, 1H), 10.47 (s, 1H), 7.88 (dd, J=13.3, 7.5Hz, 1H), 7.58 (d, J=3.6 Hz, 1H), 7.54-7.35 (m, 3H), 3.62 (s, 3H), 2.41(s, 3H), 2.19 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd for C₁₉H₁₈F₂N₃O₃:419.4, found: 419.4.

4-(2-((cyanomethyl)amino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid (80)

To a solution of2-(5-ethoxycarbonyl-1,3,5-trimethyl-pyrrol-3-yl)-2-oxo-acetic acid 4(3.0 g, 11.8 mmol) in DMF (20 mL) and CH₂Cl₂ (20 mL) was added1,1′-carbonyldiimidazole (2.3 g, 14.2 mmol) and 2-aminoacetonitrilehydrochloride (1.63 g, 17.8 mmol) and diisopropylethylamine (4.12 mL,23.7 mmol). After stirring for 2 h at room temperature, the reactionmixture was poured onto a saturated solution of ammonium chloride andextracted with CH₂Cl₂ (3×100 mL). The combined organic layers were driedover sodium sulfate and concentrated in vacuo to give 79 as a solid. Tothe crude ethyl4-(2-((cyanomethyl)amino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylate79 dissolved in methanol (10 mL) and THF (10 mL) was added a 5% solutionof sodium hydroxide (10 mL). The reaction mixture was stirred at roomtemperature overnight and after evaporation of the methanol and THF invacuo, the aqueous solution was washed with ethyl acetate (2×50 mL),acidified with a 1N HCl solution (pH=1) and extracted again with ethylacetate (3×50 mL). The combined organic layers were dried over sodiumsulfate and concentrated in vacuo. The resulting solid was washed withdiethyl ether (50 mL) and hexanes (50 mL) to yield4-(2-((cyanomethyl)amino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid 80 (0.41 g, 15.5 mmol, 13%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.79 (s,1H), 9.46 (s, 1H), 4.32 (d, J=5.7 Hz, 2H), 3.76 (s, 3H), 2.37 (s, 3H),2.36 (s, 3H). MS (ESI): m/z [M+H]⁺ calcd for C₁₂H₁₄N₃O₄: 264.3, found:264.4.

4-(2-((cyanomethyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(81)

4-(2-((cyanomethyl)amino)-2-oxoacetyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxylicacid 80 (2 g, 7.6 mmol), 3,4-difluoroaniline (1.47 g, 11.4 mmol) andDIPEA (1.98 mL, 11.4 mmol) in DMF (30 mL) was added HATU (3.18 g, 8.3mmol) at room temperature. The mixture was stirred at 65° C. for 18 h.The reaction mixture was then poured into a saturated solution ofammonium chloride and extracted with ethyl acetate (3×150 mL). Thecombined organic layers were dried over sodium sulfate and concentratedin vacuo. The residue was purified by flash chromatography(Hexanes:EtOAc=6:4 v/v) to give compound 81 as a white powder (30%, 840mg, 2.2 mmol). ¹H NMR (400 MHz, Acetone-d₆) δ 9.52 (s, 1H), 8.52 (s,1H), 8.09-7.90 (m, 1H), 7.58-7.45 (m, 1H), 7.40-7.23 (m, 1H), 4.49-4.36(m, 2H), 3.74-3.65 (m, 3H), 2.46-2.40 (m, 3H), 2.34-2.25 (m, 3H). MS(ESI): m/z [M+H]⁺ calcd for C₁₈H₁₇F₂N₄O₃: 375.3, found: 375.4.

4-(2-(((2H-tetrazol-5-yl)methyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide(82)

Sodium azide (26 mg, 0.4 mmol) and zinc bromide (90 mg, 0.4 mmol) wereadded to a suspension of4-(2-((cyanomethyl)amino)-2-oxoacetyl)-N-(3,4-difluorophenyl)-1,3,5-trimethyl-1H-pyrrole-2-carboxamide81 (50 mg, 0.1 mmol) in isopropanol (2 mL). The mixture was heated to110° C. for 20 min under microwave irradiations. The reaction mixturewas then poured onto a saturated solution of sodium carbonate (50 mL)and washed with ethyl acetate (3×20 mL). The aqueous phase was thenacidified to pH˜1 and extracted with ethyl acetate (3×50 mL). Thecombined organic phases were dried over sodium sulfate and concentratedin vacuo. The resulting residue was purified by flash chromatography(DCM/MeOH=95:5 v/v), resulting in compound 82 as a white powder (48%, 27mg, 0.1 mmol). ¹H NMR (400 MHz, DMSO-d₅) δ 10.41 (s, 1H), 9.46 (t, J=5.8Hz, 1H), 7.92-7.83 (m, 1H), 7.50-7.37 (m, 2H), 4.69 (d, J=5.7 Hz, 2H),3.58 (s, 3H), 2.33 (s, 3H), 2.17 (s, 3H). LCMS (ESI): m/z [M+H]⁺ calcdfor C₁₈H₁₈F₂N₇O₃: 418.4, found: 418.4.

N-(3,4-difluorophenyl)-1,3,5-trimethyl-4-(2-((3-(1-methyl-1H-imidazol-2-yl)prop-2-yn-1-yl)amino)-2-oxoacetyl)-1H-pyrrole-2-carboxamide(83)

Compound 7a (100 mg, 268 μmol), 2-bromo-1-methyl-1H-imidazole (65 mg,402 μmol), triethylamine (73 μL, 541 μmol), and dimethylformamide (2 mL)were combined in a sealed tube. The mixture was sparged for 2 minuteswith nitrogen, and bis(triphenylphosphine) palladium dichloride (19 mg,27 μmop was added followed by copper iodide (10 mg, 53 μmop. The mixturewas sparged again with nitrogen and stirred for 20 minutes at 70° C.under microwave irradiation. The reaction was diluted with ethyl acetate(50 mL), washed with a saturated solution of ammonium chloride (50 mL).The organic phase was dried over sodium sulfate and concentrated underreduced pressure and purified by flash chromatography on silica gel(DCM/MeOH: 96/4) to give compound 83 (62 mg, 137 μmol, 51%). ¹H NMR (400MHz, Acetone-d₆) δ 9.51 (s, 1H), 8.35 (s, 1H), 7.99 (ddd, J=13.2, 7.4,2.6 Hz, 1H), 7.57-7.48 (m, 1H), 7.34 (dt, J=10.5, 9.0 Hz, 1H), 7.14 (d,J=1.2 Hz, 1H), 6.92 (d, J=1.2 Hz, 1H), 4.46 (d, J=5.8 Hz, 2H), 3.75 (s,3H), 3.70 (s, 3H), 2.46 (s, 3H), 2.32 (s, 3H). ¹⁹F NMR (377 MHz,Acetone-d₆) δ −139.91-−140.01 (dt, J=22.1, 11.6 Hz), −147.12-−147.23(dt, J=20.5, 10.4 Hz). MS (ESI): m/z [M+H]⁺ calcd for C₂₃H₂₂F₂N₅O₃:454.4, found: 454.4.

Example 2

Cellular Toxicity Assays

The toxicity of the compounds was assessed in Vero, human PBM, CEM(human lymphoblastoid), MT-2, and HepG2 cells, as described previously(see Schinazi R. F., Sommadossi J.-P., Saalmann V., Cannon D. L., XieM.-Y., Hart G. C., Smith G. A. & Hahn E. F. Antimicrob. AgentsChemother. 1990, 34, 1061-67). Cycloheximide was included as positivecytotoxic control, and untreated cells exposed to solvent were includedas negative controls. The cytotoxicity IC₅₀ was obtained from theconcentration-response curve using the median effective method describedpreviously (see Chou T.-C. & Talalay P. Adv. Enzyme Regul. 1984, 22,27-55; Belen'kii M. S. & Schinazi R. F. Antiviral Res. 1994, 25, 1-11).The results are shown in Table 1 below:

TABLE 1 Cytotoxicity, CC₅₀, μM (% inhibition) Cytotoxicity; CC₅₀ (μM)Compound PBM CEM VERO HepG2  7a >100 >100 >90 17 >100 53 41 13 64.8 17.566.0  8 60.5 45.4 >100 98.6 36 13.9 36.7 22.9 43.5 13 >100 78.355.6 >100 20 >100 >100 >100 >100 31 >100 >100 41.9 93.9 27 37.980.1 >100 28 >100 13.4 >100 29 >100 62.4 >100 30 >100 91.4 >10025 >100 >100 68.1 >100 26 >100 >100 31.2 >100 32 >100 >100 >100 >100 7b >100 >100 >100 >100 38 >100 29 45 85 43 >100 50 >100 >100 44 9217 >100 90 45 90 36 47 75 46 >100 38 10 7 47 >100 47 51 35 48 >10027 >100 84 52 (3057) >100 50 >100 >100 54 3 4 6 55 >100 38 15 8856 >100 >100 >100 >100 57 >100 >100 >100 >100 58 >100 >100 >100 >100 5944 3 12 43 60 86 23 82 >100 61 >100 >100 >100 >10062 >100 >100 >100 >100 63 >100 >100 >100 >100 64 >100 18 96 >10065 >100 >100 >100 >100 66 >100 52 70 83 69 >100 >100 48 59 72 30 14 51100 75 >100 66 33 >100 78 >100 15 >100 84 81 30 33 >100 7182 >100 >100 >100 >100 83 7 4 7 42

Example 3

Mitochondrial Toxicity Assays in HepG2 Cells:

i) Effect of Compounds on Cell Growth and Lactic Acid Production:

The effect on the growth of HepG2 cells was determined by incubatingcells in the presence of 0 μM, 0.1 μM, 1 μM, 10 μM and 100 μM drug.Cells (5×10⁴ per well) were plated into 12-well cell culture clusters inminimum essential medium with nonessential amino acids supplemented with10% fetal bovine serum, 1% sodium pyruvate, and 1%penicillin/streptomycin and incubated for 4 days at 37° C. At the end ofthe incubation period the cell number was determined using ahemocytometer. Also taught by Pan-Zhou X-R, Cui L, Zhou X-J, SommadossiJ-P, Darley-Usmer V M. “Differential effects of antiretroviralnucleoside analogs on mitochondrial function in HepG2 cells,”Antimicrob. Agents Chemother. 2000; 44: 496-503.

To measure the effects of the compounds on lactic acid production, HepG2cells from a stock culture were diluted and plated in 12-well cultureplates at 2.5×10⁴ cells per well. Various concentrations (0 μM, 0.1 μM,1 μM, 10 μM and 100 μM) of compound were added, and the cultures wereincubated at 37° C. in a humidified 5% CO₂ atmosphere for 4 days. At day4, the number of cells in each well was determined and the culturemedium collected. The culture medium was then filtered, and the lacticacid content in the medium was determined using a colorimetric lacticacid assay (Sigma-Aldrich). Since lactic acid product can be considereda marker for impaired mitochondrial function, elevated levels of lacticacid production detected in cells grown in the presence of testcompounds would indicate a drug-induced cytotoxic effect.

ii) Effect on Compounds on Mitochondrial DNA Synthesis:

A real-time PCR assay to accurately quantify mitochondrial DNA contenthas been developed (see Stuyver L J, Lostia S, Adams M, Mathew J S, PaiB S, Grier J, Tharnish P M, Choi Y, Chong Y, Choo H, Chu C K, Otto M J,Schinazi R F. Antiviral activities and cellular toxicities of modified2′,3′-dideoxy-2′,3′-didehydrocytidine analogs. Antimicrob. AgentsChemother. 2002; 46: 3854-60). This assay was used in all studiesdescribed in this application that determine the effect of compounds onmitochondrial DNA content. In this assay, low-passage-number HepG2 cellswere seeded at 5,000 cells/well in collagen-coated 96-well plates. Testcompounds were added to the medium to obtain final concentrations of 0μM, 0.1 μM, 10 μM and 100 μM. On culture day 7, cellular nucleic acidswere prepared by using commercially available columns (RNeasy 96 kit;Qiagen). These kits co-purify RNA and DNA, and hence, total nucleicacids are eluted from the columns. The mitochondrial cytochrome coxidase subunit II (COXII) gene and the β-actin or rRNA gene wereamplified from 5 μl of the eluted nucleic acids using a multiplex Q-PCRprotocol with suitable primers and probes for both target and referenceamplifications. For COXII the following sense, probe and antisenseprimers were used, respectively: 5′-TGCCCGCCATCATCCTA-3′,5′-tetrachloro-6-carboxyfluorescein-TCCTCATCGCCCTCCCATCCC-TAMRA-3′ and5′-CGTCTGTTATGTAAAGGATGCGT-3′. For exon 3 of the β-actin gene (GenBankaccession number E01094) the sense, probe, and antisense primers are5′-GCGCGGCTACAGCTTCA-3′, 5′-6-FAMCACCACGGCCGAGCGGGATAMRA-3′ and5′-TCTCCTTAATGTCACGCACGAT-3′, respectively. The primers and probes forthe rRNA gene are commercially available from Applied Biosystems. Sinceequal amplification efficiencies are obtained for all genes, thecomparative CT method was used to investigate potential inhibition ofmitochondrial DNA synthesis. The comparative CT method uses arithmeticformulas in which the amount of target (COXII gene) is normalized to theamount of an endogenous reference (the β-actin or rRNA gene) and isrelative to a calibrator (a control with no drug at day 7). Thearithmetic formula for this approach is given by 2−ΔΔCT, where ΔΔCT is(CT for average target test sample CT for target control)−(CT foraverage reference test−CT for reference control) (see Johnson M R, KWang, J B Smith, M J Heslin, R B Diasio. Quantitation ofdihydropyrimidine dehydrogenase expression by real-time reversetranscription polymerase chain reaction. Anal. Biochem. 2000;278:175-184). A decrease in mitochondrial DNA content in cells grown inthe presence of drug indicated mitochondrial toxicity.

The effect of compounds 7 and 9 on the levels of mitochondrial andnuclear DNA, and lactic acid production was evaluated in HepG2 cells(14-day assay), and the data is tabulated below in Table 2:

TABLE 2 % inhibition IC₅₀, μM Lactic acid MtDNA/ MtDNA/ MtDNA productionCmpd μM nDNA nDNA (% of control) (% of control) 7a 10  <1/5.4 >50/>50 110 (100-121) 103 ± 32.0 25 <1/<1 91.8 (80.2-105) 106 ± 19.5 5017.7/13.1 94.6 (73.8-121) 193 ± 12.6 ddC 10 97.4/52.4 <10/<10  5.3(5.0-5.8) 196 ± 73.0 (control) 3TC 10  8.8/29.6 >10/>10  130 (85.7-196) 94 ± 20.8 (control)

The data show that compounds 7a, as described herein, is non-toxic up to25 μM and very low toxicity was noted even at 50 μM, similar to thenegative control 3TC.

Example 4

Mitochondrial Toxicity Assays in Neuro2A Cells

To estimate the potential of the compounds of this invention to causeneuronal toxicity, mouse Neuro2A cells (American Type Culture Collection131) can be used as a model system (see Ray A S, Hernandez-Santiago B I,Mathew J S, Murakami E, Bozeman C, Xie M Y, Dutschman G E, Gullen E,Yang Z, Hurwitz S, Cheng Y C, Chu C K, McClure H, Schinazi R F, AndersonK S. Mechanism of anti-human immunodeficiency virus activity ofbeta-D-6-cyclopropylamino-2′,3′-didehydro-2′,3′-dideoxyguanosine.Antimicrob. Agents Chemother. 2005, 49, 1994-2001). The concentrationsnecessary to inhibit cell growth by 50% (CC₅₀) can be measured using the3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide dye-basedassay, as described. Perturbations in cellular lactic acid andmitochondrial DNA levels at defined concentrations of drug can becarried out as described above. ddC and AZT can be used as controlnucleoside analogs.

Example 5

Assay for Bone Marrow Cytotoxicity

Primary human bone marrow mononuclear cells can be obtained commerciallyfrom Cambrex Bioscience (Walkersville, Md.). CFU-GM assays is carriedout using a bilayer soft agar in the presence of 50 units/mL humanrecombinant granulocyte/macrophage colony-stimulating factor, whileBFU-E assays used a ethylcellulose matrix containing 1 unit/mLerythropoietin (see Sommadossi J P, Carlisle R. Toxicity of3′-azido-3′-deoxythymidine and 9-(1,3-dihydroxy-2-propoxymethyl) guaninefor normal human hepatopoietic progenitor cells in vitro. Antimicrob.Agents Chemother. 1987; 31: 452-454; Sommadossi, J P, Schinazi, R F,Chu, C K, and Xie, M Y. Comparison of cytotoxicity of the (−) and (+)enantiomer of 2′,3′-dideoxy-3′-thiacytidine in normal human bone marrowprogenitor cells. Biochem. Pharmacol. 1992; 44:1921-1925). Eachexperiment can be performed in duplicate in cells from three differentdonors. AZT is used as a positive control. Cells can be incubated in thepresence of the compound for 14-18 days at 37° C. with 5% CO₂, andcolonies of greater than 50 cells can be counted using an invertedmicroscope to determine the IC₅₀. The 50% inhibitory concentration(IC₅₀) can be obtained by least-squares linear regression analysis ofthe logarithm of drug concentration versus BFU-E survival fractions.Statistical analysis can be performed with Student's t test forindependent non-paired samples.

Example 6

Anti-HBV Assay

The anti-HBV activity of the compounds was determined by treating theAD-38 cell line carrying wild type HBV under the control of tetracycline(see Ladner S. K., Otto M. J., Barker C. S., Zaifert K., Wang G. H., GuoJ. T., Seeger C. & King R. W. Antimicrob. Agents Chemother. 1997, 41,1715-20). Removal of tetracycline from the medium [Tet (−)] results inthe production of HBV. The levels of HBV in the culture supernatantfluids from cells treated with the compounds were compared with that ofthe untreated controls. Control cultures with tetracycline [Tet (+)]were also maintained to determine the basal levels of HBV expression.3TC was included as positive control.

The median effective concentrations (EC₅₀) ranges of several of thecompounds described herein against HBV are shown in Table 3:

A=1-9 μM B=0.1-0.9 μM C=0.01-0.09 μM D=0.001-0.009 μM E=0.0001-0.0009 μM

TABLE 3 Anti-HBV activity EC₅₀ EC₉₀  7a D C 17 B A 13 B B  8 D C 36 B A13 B A 20 B B 31 C A 27 C A 28 C A 29 B A 30 A A 25 C A 26 B A 32 A A 7b C B 38 D B 43 C B 44 B A 45 B A 46 B A 47 C B 48 D C 52 C B 54 A 55B A 56 >A 57 >A 58 C B 59 A 60 B B 61 B B 62 A A 63 B A 64 E D 65 C B 66D C 69 D C 72 B B 75 D C 78 D C 81 D B 82 A 83 A  3TC B A

Example 7

Production of secreted HBeAg is predominantly cccDNA-dependent inHepAD38 cells and therefore can serve as a surrogate marker for cccDNA(Ladner, S. K., Otto, M. J., Barker, C. S., Zaifert, K., Wang, G. H.,Guo, J. T., Seeger, C., King, R. W. Antimicrob Agents Chemother 1997,41, 1715-1720; Zhou T, Guo H, Guo J T, Cuconati A, Mehta A, Block T M.Antiviral Res. 2006; 72 (2): 116-24.). The effect on the levels ofcccDNA formation was assessed using a cell-based assay that measures HBVe antigen (HBeAg) as a cccDNA-dependent marker in the HepAD38 system.HepAD38 cells were seeded at 50,000 cells/well in collagen-coated96-well plates with DMEM/F12 medium (Life Technologies) supplementedwith 10% heat-inactivated fetal bovine serum. Cells were treated with0.3 μg/ml tetracycline as needed. Test compounds and controls were addedto cells to a final concentration of 10 μM or in a dose response mannerranging from 0.001 to 10 μM. Medium and test compounds were replenishedevery 5 days in culture. Supernatants were harvested at day-14,clarified by centrifugation at 5000 rpm for 5 min, and stored at −70° C.until use. ELISA—Culture medium was diluted 1:15 in DMEM/F12 and thelevels of HBeAg secreted in the culture medium were measured by usingHBeAg ELISA kit (BioChain Institute Inc. Hayward, Calif.) according tothe manufacturer's protocol. The concentration of compound that reducedlevels of secreted HBeAg by 50% (EC₅₀) was determined by linearregression.

TABLE 4 Anti-HBeAg activity (μM) Compound EC₅₀ EC₉₀  7a 0.008 0.58 13<10 ND (86%)

Example 8

Interestingly, some of the compounds synthesized and found activeagainst HBV in vitro were also unexpectedly active against West NileVirus (WNV). WNV is a mosquito-borne zoonotic arbovirus belonging to thegenus Flavivirus in the family Flaviviridae. The genetic material of WNVis a positive-sense, single strand of RNA, which is between 11,000 and12,000 nucleotides long; these genes encode seven nonstructural proteinsand three structural proteins. The RNA strand is held within anucleocapsid formed from 12-kDa protein blocks; the capsid is containedwithin a host-derived membrane altered by two viral glycoproteins.

Antiviral Screening Using a Luciferase Reporter Replicon of West NileVirus (WNV).

Baby hamster kidney (BHK) cells containing a luciferase reporterreplicon of WNV (See Shi P Y, Tilgner M, Lo M K. Virology. 2002; 296(2): 219-33) were used for high throughput screening. Renilla luciferaseand blasticitin resistance genes as a selectable marker gene wereengineered in the replicon to replace viral structural proteins.Luciferase activities were measured after 48 h of incubation usingRenilla Luciferase Assay system (Promega).

The susceptibility of West Nile virus to the compounds described hereincan also be evaluated using the assay previously described in: Song, G.Y., Paul, V., Choo, H., Morrey, J., Sidwell, R. W., Schinazi, R. F.,Chu, C. K. Enantiomeric synthesis of D- and L-cyclopentenyl nucleosidesand their antiviral activity against HIV and West Nile virus. J. Med.Chem. 2001, 44, 3985-3993,

Example 9

The susceptibility of Yellow fever to the compounds described herein canalso be assayed as previously described in: Julander, J. G., Furuta, Y.,Shafer, K., Sidwell, R. W. Activity of T-1106 in a Hamster Model ofYellow Fever Virus Infection. Antimicrob. Agents Chemother. 2007, 51,1962-1966.

Example 10

The susceptibility of Dengue to the compounds described herein can beevaluated using the high throughput assay disclosed by Lim et al., Ascintillation proximity assay for dengue virus NS52′-O-methyltransferase—kinetic and inhibition analyses, AntiviralResearch, Volume 80, Issue 3, December 2008, Pages 360-369.

Dengue virus (DENV) NS5 possesses methyltransferase (MTase) activity atits N-terminal amino acid sequence and is responsible for formation of atype 1 cap structure, m7GpppAm2′-O in the viral genomic RNA. Optimal invitro conditions for DENV2 2′-O-MTase activity can be characterizedusing purified recombinant protein and a short biotinylated GTP-cappedRNA template. Steady-state kinetics parameters derived from initialvelocities can be used to establish a robust scintillation proximityassay for compound testing. Pre-incubation studies by Lim et al.,Antiviral Research, Volume 80, Issue 3, December 2008, Pages 360-369,showed that MTase-AdoMet and MTase-RNA complexes were equallycatalytically competent and the enzyme supports a random bi bi kineticmechanism. Lim validated the assay with competitive inhibitory agents,S-adenosyl-homocysteine and two homologues, sinefungin anddehydrosinefungin. A GTP-binding pocket present at the N-terminal ofDENV2 MTase was previously postulated to be the cap-binding site. Thisassay allows rapid and highly sensitive detection of 2′-O-MTase activityand can be readily adapted for high-throughput screening for inhibitorycompounds. It is also suitable for determination of enzymatic activitiesof a wide variety of RNA capping MTases.

Example 11 Anti-Norovirus Activity

Compounds can exhibit anti-norovirus activity by inhibiting noroviruspolymerase and/or helicase, by inhibiting other enzymes needed in thereplication cycle, or by other pathways.

There is currently no approved pharmaceutical treatment for Norovirusinfection, and this has probably at least in part been due to the lackof availability of a cell culture system. Recently, a replicon systemhas been developed for the original Norwalk G-I strain (Chang, K. O., etal. (2006) Virology 353:463-473).

Both Norovirus replicons and Hepatitis C replicons require viralhelicase, protease, and polymerase to be functional in order forreplication of the replicon to occur. Most recently, an in vitro cellculture infectivity assay has been reported utilizing Norovirusgenogroup I and II inoculums (Straub, T. M. et al. (2007) Emerg. Infect.Dis. 13(3):396-403). This assay is performed in a rotating-wallbioreactor utilizing small intestinal epithelial cells on microcarrierbeads. The infectivity assay can be used to screen entry inhibitors.

Example 12

Anti-Chikungunya Activity

Anti-Chikungunya Activity can be evaluated as outlined in“Anti-Chikungunya Viral Activities of Aplysiatoxin-Related Compoundsfrom the Marine Cyanobacterium Trichodesmium erythraeum” Gupta, D. K.;Kaur, P.; Leong, S. T.; Tan, L. T.; Prinsep, M. R.; Chu, J J. H. MarDrugs. January 2014; 12(1): 115-127; 10.3390/md12010115 and referencescited therein.

Example 13

Anti-HCV Activity

The anti-HCV activity of the compounds described herein can be measured,for example, using an HCV replicon assay as described, for example, inStuyver L et al., Ribonucleoside analogue that blocks replication orbovine viral diarrhea and hepatitis C viruses in culture. Antimicrob.Agents Chemother. 2003, 47, 244-254.

In this assay, Huh 7 Clone B cells containing HCV Replicon RNA can beseeded in a 96-well plate at 5000 cells/well, and the compounds testedat 10 μM in triplicate immediately after seeding. Following five daysincubation (37° C., 5% CO2), total cellular RNA can be isolated by usinga versaGene RNA purification kit from Gentra. Replicon RNA and aninternal control (TaqMan rRNA control reagents, Applied Biosystems) canbe amplified in a single step multiplex Real Time RT-PCR Assay. Theantiviral effectiveness of the compounds can be calculated bysubtracting the threshold RT-PCR cycle of the test compound from thethreshold RT-PCR cycle of the no-drug control (ΔCt HCV). A ΔCt of 3.3equals a 1-log reduction (equal to 90% less starting material) inReplicon RNA levels. The cytotoxicity of the compounds can also becalculated by using the ΔCt rRNA values. 2′-C-Me-C can be used as thepositive control. To determine EC₉₀ and IC₅₀ values, ΔCt: values canfirst be converted into fraction of starting material and then used tocalculate the % inhibition.

To look specifically at whether the compounds inhibit HCV NS5B, one canusing an assay such as that described in “A complex network ofinteractions between S282 and G283 of HCV NS5B and the template strandaffect susceptibility to Sofosbuvir and Ribavirin,” Kulkarni et al.,Antimicrob Agents Chemother. 2016 Jan. 11. pii: AAC.02436-15.

Example 14

Capsid Formation Assay for Use in Monitoring HBV Capsid Assembly

In the absence of compounds which disrupt capsid formation, hepatitis Bvirus core C-terminally truncated protein (HBV Cp149, protein isolatedby reported methods [Zlotnick, A et al; Biochem 1996, 35, 7412-7421]normally assembles into an HBV Cp149 capsid. The purpose of this examplewas to determine whether putative active agents would disrupt capsidformation, and thus be active as anti-HBV agents. Putative active agentswere incubated at a concentration of 25 μM for 1 h at 4° C. with HBVCp149 at a concentration of 10 μM. Capsid assembly was then promoted byadding 300 mM NaCl, and storing the mixture overnight at 4°.Negative-stain electron micrographs were collected using a JEOL JEM-1400120 kV electron microscope using uranyl acetate as a contrast agent.These images showed whether capsids formed, and if they did form,whether they formed fully-formed hollow spheres, or deformed (i.e., forexample, misassembled or incomplete) spheres.

When treated with vehicle, the capsid formation proceed as expected,forming fully-formed hollow spheres with a diameter of approximately 40nm. When compound GLS4 was added, the capsid formation was disrupted, asshown by the formation of relatively large (around 80-100 nm)misassembled hollow spheres. When Compound 7a was added, the capsidformation was disrupted, as shown by the formation of relatively small(less than around 40 nm) and tightly packed incomplete hollow spheres.The results are shown in FIG. 1.

The next question was whether these compounds could disruptalready-formed capsids. Accordingly, the capsids were formed asdiscussed above (incubation of isolated HBV Cp149 with 300 mM NaCl,stored overnight at 4°). Then, the capsids were incubated with theputative compound (overnight at 4°), and electron micrographs were thentaken. FIG. 2 shows electron micrographs of capsids incubated withvehicle, with 25 μM GLS4, and with 25 μM Compound 7a. FIG. 3 shows theresults with GLS4, in which the capsids were disrupted. The micrographsshow that the capsids were broken, like cracked egg shells. FIG. 4 showsthe results with Compound 7a, where the concentration of capsids wasclearly, and significantly reduced, and the remaining capsids wererelatively small and tightly packed.

Various publications are cited herein, the disclosures of which areincorporated by reference in their entireties for all purposes.

The present invention is not to be limited in scope by the specificembodiments described herein. Indeed, various modifications of theinvention in addition to those described will become apparent to thoseskilled in the art from the foregoing description and accompanyingfigures. Such modifications are intended to fall within the scope of theappended claims.

1. A compound of the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein whenR¹ and R^(1′) are attached to a carbon, they are, independently,hydrogen, halogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonyl alkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl; when R¹ and R^(1′) are attached to a nitrogen,they are, independently, hydrogen, C₂₋₆ alkoxy, C₃₋₆ alkoxyalkyl, C₂₋₆alkenyl, alkoxycarbonyl, carbonylalkyl, carbonyl aryl, C₁₋₆ alkyl,heterocyclylalkyl, C₂₋₆ hydroxyalkyl, or S(O)₂R′; each R′ isindependently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₂₋₆ alkenyl,C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl, alkylaryl, orarylalkyl, or if two R′ reside on the same nitrogen atom, they can cometogether to form a C₃₋₆ ring optionally containing a N, O, or Sheteroatom; the R′ groups can optionally be substituted with one or moresubstituents, which substituents are, independently, halo, C₁₋₆haloalkyl, C₁₋₆ hydroxyalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy,amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, alkoxyalkyl, aryloxy, nitro, cyano, sulfonic acid, thiol, imine,sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid, orphosphonate; u and v are independently 0, 1, 2, 3, 4 or 5; I is phenyl,a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms, which are, independently, N, O, or S, a C₄₋₁₄bicyclic ring; alkylheteroaryl, or alkylaryl; J is a five-memberedheteroaromatic ring containing one, two, or three heteroatoms, whichare, independently, N, O, or S, a six or seven-membered ring or a six orseven-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S; or a four-membered ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S, W is

R¹² is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, R¹³is C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl, alkylaryl, arylalkyl, aC₄₋₁₄ bicyclic ring; or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, R¹³ is optionally substituted with one or more substituents eachindependently selected from the group consisting of hydrogen, halogen,CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl,arylalkoxycarbonyl, carboxyl, haloalkyl, heterocyclylalkyl, C₁₋₆hydroxyalkyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl, where substituents on the substituted aryl and substitutedheteroaryl are selected from the group consisting of halogen, SF₅, CF₃,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano,azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, and C₁₋₆ alkyl. or R¹² and R¹³ together with thenitrogen to which they are attached form a 3 to 4 membered ringoptionally substituted with one or more substituents each independentlyselected from the group consisting of hydrogen, halogen (F, Cl, Br, I),CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano,azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, and C₁₋₆ hydroxyalkyl; or of the followingformula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein: A isselected from the group consisting of phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or three nitrogenatoms which are, independently, N, O, or S; a C₄₋₁₄ bicyclic ring,alkylheteroaryl, and alkylaryl; B is a six or seven-membered ring or asix or seven-membered bridged or spiro-fused ring containing zero, one,or two heteroatoms, which are, independently, N, O, or S, afive-membered ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S; a four-membered ring containing zero, one, ortwo heteroatoms, which are, independently, N, O, or S, or a C₄₋₁₄bicyclic ring, when R¹ and R^(1′) are attached to a carbon, they are,independently, hydrogen, halogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonyl alkyl,C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; when R¹ and R^(1′) are attachedto a nitrogen, they are, independently, hydrogen, C₂₋₆ alkoxy, C₃₋₆alkoxyalkyl, C₂₋₆ alkenyl, alkoxycarbonyl, carbonylalkyl, carbonyl aryl,C₁₋₆ alkyl, heterocyclylalkyl, C₂₋₆ hydroxyalkyl, or S(O)₂R′; each R′ isindependently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₂₋₆ alkenyl,C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl, alkylaryl, orarylalkyl, or if two R′ reside on the same nitrogen atom, they can cometogether to form a C₃₋₆ ring optionally containing a N, O, or Sheteroatom; the R′ groups can optionally be substituted with one or moresubstituents, which substituents are, independently, halo, C₁₋₆haloalkyl, C₁₋₆ hydroxyalkyl, hydroxyl, carboxyl, acyl, aryl, acyloxy,amino, amido, carboxyl derivatives, alkylamino, dialkylamino, arylamino,alkoxy, alkoxyalkyl, aryloxy, nitro, cyano, sulfonic acid, thiol, imine,sulfonyl, sulfanyl, sulfinyl, sulfamonyl, ester, carboxylic acid, amide,phosphonyl, phosphinyl, phosphoryl, phosphine, thioester, thioether,acid halide, anhydride, oxime, hydrozine, carbamate, phosphonic acid, orphosphonate; u and v are independently 0, 1, 2, 3, 4 or 5; X is

R³ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, R²is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, heteroaryl, alkylaryl, arylalkyl, a six-membered ring ora six-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatomswhich are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms which are,independently, N, O, or S; cycloalkyl, alkylheteroaryl, or alkylaryl; R²is optionally substituted with one or more substituents, which each,independently, are halogen, CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl,C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; or is optionally substituted with aryl, substituted aryl,heteroaryl, or substituted heteroaryl, where substituents on thesubstituted aryl and substituted heteroaryl are selected from the groupconsisting of halogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl, and R²and R³ can come together with the nitrogen to which they are attached toform a 6-10 membered bicyclic or bridged ring, a 3 to 8 saturated ring,or a 5 membered unsaturated ring; such bicyclic, bridged, saturated andunsaturated rings optionally containing one or more additionalheteroatoms, where each is, independently, O, S or N, and optionallybeing substituted with one or more substituents, wherein each,independently, is halogen (including F, Cl, Br, I), CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl; or of the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein R¹ andR^(1′) are as defined with respect to Formula I, u and v areindependently 0, 1, 2, 3, 4 or 5; C is phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, or S; a C₄₋₁₄ bicyclic ring,alkylaryl, or alkylheteroaryl; D is phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, or a C₄₋₁₄ bicyclicring, Y is

R⁴ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl; R⁵ isalkylaryl, arylalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, heteroaryl, or asix-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; R⁵ is optionallysubstituted with one or more substituents, each of which is,independently, halogen, CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl,C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or is optionally substitutedwith aryl, substituted aryl, heteroaryl, or substituted heteroaryl,where substituents on the substituted aryl and substituted heteroarylare selected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl, wherein, if C is phenyl, D is not phenyl or a 5-memberedring heteroaryl, or if C is phenyl and D is phenyl or a 5-membered ringheteroaryl, then R⁵ is not alkylaryl, alkenyl, or a six-membered bridgedring; or when Y is

R⁴ and R⁵ together with the nitrogen to which they are attached form a 3to 4 membered ring optionally substituted with one or more substituents,each of which is, independently, halogen, CF₃, hydroxy, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′), C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; or of the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein R¹ andR^(1′) are as defined with respect to Formula I, u and v areindependently 0, 1, 2, 3, 4 or 5; E is a six-membered heteroaromaticring containing one, two, or three nitrogen atoms, a five-memberedheteroaromatic ring containing one, two, or three heteroatoms, whereeach is, independently, N, O, or S; a C₄₋₁₄ bicyclic ring,alkylheteroaryl, or alkylaryl; F is a five-membered heteroaromatic ringcontaining one, two, or three heteroatoms which are, independently, N,O, or S, or a C₄₋₁₄ bicyclic ring, Z is

R⁸ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, R⁹is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ring ora six-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms, which are independently N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S, a five-membered ring containingzero, one, or two heteroatoms, which are, independently, N, O, or S; afour-membered ring containing zero, one, or two heteroatoms, which are,independently, N, O, or S, or a three membered ring; R⁹ is optionallysubstituted with one or more substituents, each of which isindependently halogen, CF₃, SF₅, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, cyano, azido, C₂₋₆ alkynyl,C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,cycloalkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl,heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or is optionally substitutedwith aryl, substituted aryl, heteroaryl, or substituted heteroaryl,where substituents on the substituted aryl and substituted heteroarylare selected from the group consisting of halogen, SF₅, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido,C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, andC₁₋₆ alkyl, and R⁸ and R⁹ can come together with the nitrogen to whichthey are attached to form a 6-10 membered bicyclic or bridged ring or a3 to 8 saturated ring; such bicyclic, bridged and saturated ring moietyoptionally containing one or more additional heteroatoms which,independently, are O, S or N and optionally being substituted with oneor more substituents, each, independently, is halogen, CF₃, hydroxy,N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl,or C₁₋₆ hydroxyalkyl; or of the following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein G isphenyl, a six-membered heteroaromatic ring containing one, two, or threenitrogen atoms, a five-membered heteroaromatic ring containing one, two,or three heteroatoms, which are, independently, N, O, or S; a C₄₋₁₄bicyclic ring, alkylheteroaryl, or alkylaryl; H is phenyl, asix-membered heteroaromatic ring containing one, two, or three nitrogenatoms, a six-membered non-aromatic ring optionally containing one, two,or three heteroatoms, which are, independently, N, O, or S; or a C₄₋₁₄bicyclic ring; when R¹ and R^(1′) are attached to a carbon they are,independently, hydrogen, halogen, CF₃, hydroxy, SF₅, N(R′)S(O)₂R′,S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; when R¹ and R^(1′) are attached to a nitrogen they are,independently, hydrogen, C₁₋₆ alkoxy, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,carbonylalkyl, carbonyl aryl, C₁₋₆ alkyl, C₂₋₆ alkynyl, C₂₋₆ alkenyl,heterocyclylalkyl, C₁₋₆ hydroxyalkyl, or S(O)₂R′; each R′ isindependently H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₂₋₆ alkenyl,C₂₋₆ alkynyl, C₃₋₆ cycloalkyl, aryl, heteroaryl, alkylaryl, orarylalkyl, or if two R′ reside on the same nitrogen atom they can cometogether to form a C₃₋₆ alkyl ring optionally containing a N, O, or S;wherein the R′ groups can be substituted with one or more C₁₋₆hydroxyalkyl, aminoalkyl, or alkoxyalkyl substituents; u and v areindependently 0, 1, 2, 3, 4 or 5; W is

R¹⁰ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, R¹¹is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, aryl, heteroaryl, alkylaryl, arylalkyl, phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, asix-membered ring or a six-membered bridged or spiro-fused ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S, a seven-membered bridged or spiro-fused ring containing zero,one, or two heteroatoms, which are, independently, N, O, or S, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms, which are, independently, N, O, or S, a five-membered ringcontaining zero, one, or two heteroatoms, which are, independently, N,O, or S; a four-membered ring containing zero, one, or two heteroatoms,which are, independently, N, O, or S; a three membered ring,alkylheteroaryl, or alkylaryl; wherein R¹¹ is optionally substitutedwith one or more substituents selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, C₁₋₆hydroxyalkyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl, where substituents on the substituted aryl and substitutedheteroaryl are selected from the group consisting of halogen, SF₅, CF₃,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano,azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, and C₁₋₆ alkyl; and R¹⁰ and R¹¹ can come togetherwith the nitrogen to which they are attached form a 6-10 memberedbicyclic or bridged ring or a 3 to 8 saturated ring; such bicyclic,bridged or saturated ring moiety optionally containing one or moreadditional heteroatoms, which are each, independently, O, S or N, andoptionally substituted with one or more substituents, each of which is,independently, halogen, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl,carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆ hydroxyalkyl; or ofthe following formula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein R¹ andR^(1′) are as defined with respect to Formula I, u and v areindependently 0, 1, 2, 3, 4 or 5; K is a six-membered heteroaromaticring containing one, two, or three nitrogen atoms, a five-memberedheteroaromatic ring containing one, two, or three heteroatoms which are,independently, N, O, or S; a C₄₋₁₄ bicyclic ring, alkylheteroaryl, oralkylaryl; L is a five-membered heteroaromatic ring containing one, two,or three heteroatoms which are, independently, N, O, or S, a six orseven-membered ring or a six or seven-membered bridged or spiro-fusedring containing zero, one, or two heteroatoms which are, independently,N, O, or S, a five-membered ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S; a four-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,or S, or a C₄₋₁₄ bicyclic ring, W is

R¹⁴ is H, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, or C₂₋₆ alkynyl, R¹⁵is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, a six-membered ring ora six-membered bridged or spiro-fused ring containing zero, one, or twoheteroatoms which are, independently, N, O, or S, a seven-memberedbridged or spiro-fused ring containing zero, one, or two heteroatomswhich are, independently, N, O, S, or Se, a five-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,S or Se; a four-membered ring containing zero, one, or two heteroatomswhich are, independently, N, O, S, or Se; R¹⁵ is optionally substitutedwith one or more substituents which are, independently, halogen, SF₅,CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆ alkoxy, C₁₋₆haloalkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl,alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl, cycloalkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, or C₁₋₆hydroxyalkyl; or is optionally substituted with aryl, substituted aryl,heteroaryl, or substituted heteroaryl, where substituents on thesubstituted aryl and substituted heteroaryl are selected from the groupconsisting of halogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′,S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, and C₁₋₆ alkyl, or R¹⁴and R¹⁵ can come together with the nitrogen to which they are attachedform a 6-10 membered bicyclic or bridged ring or a 3 to 8 memberedsaturated ring; such bicyclic, bridged and saturated ring moietyoptionally containing one or more additional heteroatoms which are,independently, O, S or N, and optionally being substituted with one ormore substituents each independently selected from the group consistingof halogen, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆alkoxy, cyano, azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, C₁₋₆ alkyl, arylalkoxycarbonyl, carboxy, C₁₋₆haloalkyl, heterocyclylalkyl, and C₁₋₆ hydroxyalkyl; of the followingformula:

or a pharmaceutically acceptable salt or prodrug thereof, wherein R¹ andR^(1′) are as defined with respect to Formula I, u and v areindependently 0, 1, 2, 3, 4 or 5; M is phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms which are, independently, N, O, S, or Se, a C₄₋₁₄ bicyclicring, alkylheteroaryl, or alkylaryl, N is phenyl, a six-memberedheteroaromatic ring containing one, two, or three nitrogen atoms, afive-membered heteroaromatic ring containing one, two, or threeheteroatoms independently selected from the group consisting of N, O, S,and Se, a six or seven-membered ring or a six or seven-membered bridgedor spiro-fused ring containing zero, one, or two heteroatoms which are,independently, N, O, S, or Se, a five-membered ring containing zero,one, or two heteroatoms which are, independently, N, O, S, or Se; afour-membered ring containing zero, one, or two heteroatoms which are,independently, N, O, S or Se; or a C₄₋₁₄ bicyclic ring, V is

and R¹⁶ is C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₈ alkoxyalkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, aryl, heteroaryl, a six-membered ring or a six-memberedbridged or spiro-fused ring containing zero, one, or two heteroatomswhich are, independently, N, O, S, or Se, a five-membered ringcontaining zero, one, or two heteroatoms which are, independently, N, O,S, or Se; a four-membered ring containing zero, one, or two heteroatomswhich are, independently, N, O, S, or Se; alkylaryl, arylalkyl,alkylheteroaryl, or alkylaryl, wherein R¹⁶ is optionally substitutedwith one or more substituents selected from the group consisting ofhalogen, SF₅, CF₃, hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C₁₋₆alkoxy, C₁₋₆ haloalkoxy, C₂₋₆ alkenyl, cyano, C₂₋₆ alkynyl, C₃₋₆alkoxyalkyl, alkoxycarbonyl, alkoxycarbonylalkyl, C₁₋₆ alkyl,arylalkoxycarbonyl, carboxy, C₁₋₆ haloalkyl, heterocyclylalkyl, C₁₋₆hydroxyalkyl, aryl, substituted aryl, heteroaryl, and substitutedheteroaryl, where substituents on the substituted aryl and substitutedheteroaryl are selected from the group consisting of halogen, SF₅, CF₃,hydroxy, N(R′)S(O)₂R′, S(O)₂R′, S(O)₂N(R′)₂, C(O)R′, C₁₋₆ alkoxy, cyano,azido, C₂₋₆ alkynyl, C₃₋₆ alkoxyalkyl, alkoxycarbonyl,alkoxycarbonylalkyl, and C₁₋₆ alkyl.
 2. (canceled)
 3. (canceled) 4.(canceled)
 5. (canceled)
 6. (canceled)
 7. (canceled)
 8. (canceled) 9.(canceled)
 10. (canceled)
 11. A compound selected from the groupconsisting of:

and pharmaceutically acceptable salts or prodrugs thereof.
 12. Thecompound of claim 11, wherein the compound is:

and pharmaceutically acceptable salts or prodrugs thereof. 13.(canceled)
 14. (canceled)
 15. (canceled)
 16. (canceled)
 17. (canceled)18. (canceled)
 19. (canceled)
 20. (canceled)
 21. (canceled)
 22. Apharmaceutical composition comprising a compound claim 1, and apharmaceutically-acceptable carrier.
 23. (canceled)
 24. (canceled) 25.(canceled)
 26. (canceled)
 27. A method for treating a host infected withHBV, preventing an HBV infection, during an HBV infection, or reducingthe biological activity of an infection with HBV in host, comprisingadministering an effective amount of claim 1 to the host in need oftreatment thereof.
 28. The method of claim 27, wherein the methodfurther comprising administering another HBV virus agent in combinationor alternation with the compound of claim
 1. 29. (canceled)
 30. A methodfor treating a host infected with the West Nile Virus infection,preventing a West Nile Virus infection, or reducing the biologicalactivity of an infection with West Nile Virus in a host, comprisingadministering an effective amount of a compound of claim 1 to the hostin need of treatment thereof.
 31. (canceled)
 32. A method for treating ahost infected with a flaviviridae virus, preventing an infection by oneor these viruses, or reducing the biological activity of an infectionwith one of these viruses in a host, comprising administering aneffective amount of a compound of claim 1 to the host in need oftreatment thereof.
 33. (canceled)
 34. (canceled)
 35. The method of claim27, wherein the treating a host infected with HBV, preventing an HBVinfection, or curing an HBV infection suppresses HDV infection. 36.(canceled)
 37. The compound of claim 12 is:

and pharmaceutically acceptable salts and prodrugs thereof.
 38. Thecompound of claim 12 is:

and pharmaceutically acceptable salts and prodrugs thereof.
 39. Thecompound of claim 12 is:

and pharmaceutically acceptable salts and prodrugs thereof.
 40. Thecompound of claim 1, wherein R¹² is hydrogen.
 41. The compound of claim1, wherein R¹³ is C₂₋₆ alkenyl, C₂₋₆ alkynyl, aryl, or heteroaryl. 42.The compound of claim 41, wherein R¹³ is C₂₋₆ alkenyl.
 43. The compoundof claim 41, wherein R¹³ is C₂₋₆ alkynyl.
 44. The compound of claim 41,wherein R¹³ is aryl.
 45. The compound of claim 41, wherein R¹³ isheteroaryl.
 46. The compound of claim 1, wherein J is a five-memberedheteroaromatic ring containing one, two, or three heteroatoms.
 47. Thecompound of claim 46, wherein J is pyrrolyl.
 48. The compound of claim1, wherein I is phenyl.
 49. The compound of claim 1, wherein u is
 3. 50.The compound of claim 1, wherein R¹ is C₁₋₆ alkyl.
 51. The compound ofclaim 1, wherein v is
 2. 52. The compound of claim 1, wherein R^(1′) ishalogen.